Melvin G Rosenfeld, Ph.D.

New York University School of Medicine
Abramson, Steven B; Rosenfeld, Mel
2010 Sep;85(9 Suppl):S380-S386, Academic medicine
— id: 141615, year: 2010, vol: 85, page: S380, stat: Journal Article,

Neuronal molecular mimicry in immune-mediated neurologic disease
Levin, M C; Krichavsky, M; Berk, J; Foley, S; Rosenfeld, M; Dalmau, J; Chang, G; Posner, J B; Jacobson, S
1998 Jul;44(1):87-98, Annals of neurology
Molecular mimicry is implicated in the pathogenesis of autoimmune diseases such as diabetes mellitus, rheumatoid arthritis, and multiple sclerosis (MS). Cellular and antibody-mediated immune responses to shared viral-host antigens have been associated with the development of disease in these patients. Patients infected with human T-lymphotropic virus type I (HTLV-I) develop HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), an immune-mediated disorder of the central nervous system (CNS) that resembles some forms of MS. Damage to neuronal processes in the CNS of HAM/TSP patients is associated with an activated cellular and antibody-mediated immune response. In this study, IgG isolated from HAM/TSP patients was immunoreactive with uninfected neurons and this reactivity was HTLV-I specific. HAM/TSP IgG stained uninfected neurons in human CNS and cell lines but not nonneuronal cells. Neuronal western blots showed IgG reactivity with a single 33-kd band in all HAM/TSP patients tested. By contrast, no neuron-specific IgG reactivity could be demonstrated from HTLV-I seronegative controls and, more important, from HTLV-I seropositive, neurologically asymptomatic individuals. Both immunocytochemical staining and western blot reactivity were abolished by preincubating HAM/TSP IgG with HTLV-I protein lysate but not by control proteins. Staining of CNS tissue by a monoclonal antibody to HTLV-I tax (an immunodominant HTLV-I antigen) mimicked HAM/TSP IgG immunoreactivity. There was no staining by control antibodies. Absorption of HAM/TSP IgG with recombinant HTLV-I tax protein or preincubation of CNS tissue with the monoclonal antibody to HTLV-I tax abrogated the immunocytochemical and western blot reactivity of HAM/TSP IgG. Furthermore, in situ human IgG localized to neurons in HAM/TSP brain but not in normal brain. These data indicate that HAM/TSP patients develop an antibody response that targets uninfected neurons, yet reactivity is blocked by HTLV-I, suggesting viral-specific autoimmune reactivity to the CNS, the damaged target organ in HAM/TSP
— id: 125456, year: 1998, vol: 44, page: 87, stat: Journal Article,

CHARACTERIZATION OF RAB BINDING-PROTEINS THAT MAY MEDIATE THE ACTIONS OF RAB11 AND RAB8
REN, M; ZENG, J; GRAVOTTA, D; MORIMOTO, T; ROSENFELD, M; DELEMOSCHIARANDINI, C; TEMPST, P; ERDJUMENTBROMAGE, H; LUI, M; ADESNIK, M; SABATINI, DD
1995 NOV ;6(23):690-690, Molecular biology of the cell
— id: 52664, year: 1995, vol: 6, page: 690, stat: Journal Article,

Integrin alpha v beta 3 antagonists promote tumor regression by inducing apoptosis of angiogenic blood vessels
Brooks PC; Montgomery AM; Rosenfeld M; Reisfeld RA; Hu T; Klier G; Cheresh DA
1994 Dec 30;79(7):1157-1164, Cell
A single intravascular injection of a cyclic peptide or monoclonal antibody antagonist of integrin alpha v beta 3 disrupts ongoing angiogenesis on the chick chorioallantoic membrane (CAM). This leads to the rapid regression of histologically distinct human tumors transplanted onto the CAM. Induction of angiogenesis by a tumor or cytokine promotes vascular cell entry into the cell cycle and expression of integrin alpha v beta 3. After angiogenesis is initiated, antagonists of this integrin induce apoptosis of the proliferative angiogenic vascular cells, leaving preexisting quiescent blood vessels unaffected. We demonstrate therefore that ligation of integrin alpha v beta 3 is required for the survival and maturation of newly forming blood vessels, an event essential for the proliferation of tumors
— id: 34926, year: 1994, vol: 79, page: 1157, stat: Journal Article,

Carboxyl methylation of Ras-related proteins during signal transduction in neutrophils
Philips MR; Pillinger MH; Staud R; Volker C; Rosenfeld MG; Weissmann G; Stock JB
1993 Feb 12;259(5097):977-980, Science
In human neutrophils, as in other cell types, Ras-related guanosine triphosphate-binding proteins are directed toward their regulatory targets in membranes by a series of posttranslational modifications that include methyl esterification of a carboxyl-terminal prenylcysteine residue. In intact cells and in a reconstituted in vitro system, the amount of carboxyl methylation of Ras-related proteins increased in response to the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (FMLP). Activation of Ras-related proteins by guanosine-5'-O-(3-thiotriphosphate) had a similar effect and induced translocation of p22rac2 from cytosol to plasma membrane. Inhibitors of prenylcysteine carboxyl methylation effectively blocked neutrophil responses to FMLP. These findings suggest a direct link between receptor-mediated signal transduction and the carboxyl methylation of Ras-related proteins
— id: 13250, year: 1993, vol: 259, page: 977, stat: Journal Article,

Synthesis of mannosylglucosaminylinositol phospholipids in normal but not paroxysmal nocturnal hemoglobinuria cells
Hirose, S; Ravi, L; Prince, G M; Rosenfeld, M G; Silber, R; Andresen, S W; Hazra, S V; Medof, M E
1992 Jul 1;89(13):6025-6029, Proceedings of the National Academy of Sciences of the United States of America
To identify mannosyl (Man)-containing intermediates of the human glycoinositol phospholipid (GPI) anchor pathway and examine their expression in paroxysmal nocturnal hemoglobinuria (PNH), mannolipid products deriving from in vitro guanosine diphosphate [3H]Man labeling of HeLa cell microsomes were characterized. The defined GPI species were correlated with products deriving from in vivo [3H]Man labeling of normal and (GPI-anchor defective) affected leukocytes. In vitro analyses in HeLa cells showed dolichol-phosphoryl (Dol-P)-[3H]Man and a spectrum of [3H]Man lipids exhibiting TLC mobilities approximating those of Trypanosoma brucei (Tryp) GPI precursors. Iatrobead HPLC separations and partial characterizations of the major isolated [3H]Man species (designated H1-H8) showed that all but H1 (Dol-P-Man) were sensitive to HNO2 deamination and serum GPI-specific phospholipase D digestion but were resistant to phosphatidylinositol-specific phospholipase C digestion unless previously deacylated with mild alkali. [3H]Man label in H3, H4, and H6 but not in H5 or H7 was efficiently released into the aqueous phase by jack bean alpha-mannosidase digestion. BioGel P-4 and AX-5 sizing of the dephosphorylated core glycan fragments of H6 and H7 gave values that coincided precisely with the corresponding glycan fragments from the fully assembled Tryp anchor donor A' (P2). Affected leukocytes from four patients with PNH supported formation of GlcNAc- and GlcN-PI but all failed to express H6 and H7 as well as H8 and two showed complete absence of earlier Man-containing intermediates. These findings argue that human intracellular GPI mannolipids are built on acylated inositol phospholipids, that H6 and H7 contain differentially phosphoethanolamine-substituted Man3-GlcN-inositol cores, and that PNH cells are defective in conversion of GlcN-PI into these more mature mannolipid structures
— id: 135351, year: 1992, vol: 89, page: 6025, stat: Journal Article,

Glucose deprivation induces the selective accumulation of hexose transporter protein GLUT-1 in the plasma membrane of normal rat kidney cells
Haspel HC; Mynarcik DC; Ortiz PA; Honkanen RA; Rosenfeld MG
1991 Jan;5(1):61-72, Molecular endocrinology
Antibody to the carboxyl-terminal of hexose transporter protein GLUT-1 was used to localize this carrier in normal rat kidney (NRK) cells during D-glucose (Glc) deprivation. Glc-deprivation of NRK cells induces increased hexose transport, inhibits the glycosylation of GLUT-1, and increases the content of both native, 55,000 apparent mol wt (Mr) and aglyco, 38,000 Mr GLUT-1 polypeptides. The distribution of GLUT-1 protein in subcellular fractions isolated from Glc-fed NRK cells shows that the 55,000 Mr polypeptide is most abundant in intracellular membrane fractions. Glc-fed cells that have been tunicamycin treated contain principally the 38,000 Mr GLUT-1 polypeptide, which is found predominantly in intracellular membrane fractions. In Glc-deprived cells the 55,000 Mr GLUT-1 polypeptide localizes predominantly in the Golgi and plasma membrane fractions, whereas the more abundant 38,000 Mr GLUT-1 polypeptide is distributed throughout all membrane fractions. In Glc-deprived but fructose-fed cells only the 55,000 Mr GLUT-1 polypeptide is detected, and it is found predominantly in the plasma membrane and Golgi fractions. The localization of GLUT-1 protein was directly and specifically visualized in NRK cells by immunofluorescence microscopy. Glc-fed cells show little labeling of cell borders and a small punctate juxtanuclear pattern suggestive of localization to the Golgi and, perhaps, endoplasmic reticulum. Glc-fed cells that have been tunicamycin treated show large punctate intracellular accumulations suggestive of localization to distended Golgi and perhaps endoplasmic reticulum. Glc-deprived cells exhibited intense labeling of cell borders as well as intracellular accumulations. Glc-deprived but fructose-fed cells show fewer intracellular accumulations, and the labeling is, in general, limited to the cell borders. Our results suggest that Glc deprivation induces the selective accumulation of GLUT-1 in the plasma membrane of NRK cells
— id: 57474, year: 1991, vol: 5, page: 61, stat: Journal Article,

A report of familial carotid body tumors and multiple extra-adrenal pheochromocytomas
Jensen JC; Choyke PL; Rosenfeld M; Pass HI; Keiser H; White B; Travis W; Linehan WM
1991 May;145(5):1040-1042, Journal of urology
A case of familial carotid body tumors and multiple extra-adrenal pheochromocytomas is reported. The carotid body tumors, resected previously, were bilateral and associated with 4 intra-abdominal extra-adrenal pheochromocytomas. Magnetic resonance imaging was far superior to computerized tomography and 131iodine-metaiodobenzylguanidine in visualizing the intra-abdominal lesions, and may soon become the imaging technique of choice in the evaluation of patients with suspected pheochromocytoma
— id: 59172, year: 1991, vol: 145, page: 1040, stat: Journal Article,

Low molecular weight GTP-binding proteins in human neutrophil granule membranes
Philips MR; Abramson SB; Kolasinski SL; Haines KA; Weissmann G; Rosenfeld MG
1991 Jan 15;266(2):1289-1298, Journal of biological chemistry
Degranulation of neutrophils involves the differential regulation of the exocytosis of at least two populations of granules. Low molecular weight GTP-binding proteins (LMW-GBPs) have been implicated in the regulation of vesicular traffic in the secretory pathways of several types of cells. In the present study we identify distinct subsets of LMW-GBPs associated with the membranes of neutrophil-specific and azurophilic granules. Ninety-four percent of total [35S]guanosine 5'-(3-O-thio)triphosphate (GTP gamma S) binding activity was equally distributed between the plasma membrane and cytosol with the remaining 6% localized in the granules. In contrast, the cytosol contained only 10% of the total GTPase activity while the specific granules accounted for 13%. [alpha-32P]GTP binding to proteins transferred to nitrocellulose revealed LMW-GBPs in all fractions except the azurophilic granules. The specific granules contained three out of four bands which were found in the plasma membrane; these ranged from 20 to 23 kDa and all were resistant to alkaline extraction. Photoaffinity labeling with [alpha-32P]8-azido-GTP in the presence of micromolar Al3+ identified proteins of 25 and 26 kDa unique to azurophilic granules; these could not be labeled with [alpha-32P]8-azido-ATP and could be extracted by acidic but not alkaline pH. Botulinum C3-mediated [32P]ADP-ribosylation identified proteins of 16, 20, and 24 kDa both in plasma membranes and those of specific granules. An anti-ras monoclonal antibody, 142-24E5, recognized a 20-kDa protein localized to the plasma and specific granule membranes which could not be extracted by alkaline pH, was not a substrate for botulinum C3 ADP-ribosyltransferase, and was translocated from specific granules to plasma membrane after exposure of neutrophils to phorbol myristate acetate. We conclude that neutrophil-specific and azurophilic granules contain distinct subsets of LMW-GBPs which are uniquely situated to regulate the differential exocytosis of these two compartments
— id: 8314, year: 1991, vol: 266, page: 1289, stat: Journal Article,

POLYISOPRENYLATION OF RAS-RELATED PROTEINS IN HUMAN NEUTROPHILS(PMN) AND DIFFERENTIAL-EFFECTS OF LOVASTATIN (LS) AND N-ACETYL-S-FARNESYLCYSTEINE (AFC) ON SECRETION
PHILIPS, MR; ROSENFELD, MG; STOCK, JB; WEISSMANN, G
1991 APR ;39(2):A273-A273, Clinical research
— id: 51612, year: 1991, vol: 39, page: A273, stat: Journal Article,

LOCALIZATION OF LOW-MOLECULAR-WEIGHT GTP BINDING-PROTEINS IN MEMBRANES OF HUMAN NEUTROPHIL GRANULES
Philips, MR; Rosenfeld, MG; Abramson, SB; Kolasinski, SL; Haines, KA; Weissmann, G
1990 Apr;38(2):A351-A351, Clinical research
— id: 32065, year: 1990, vol: 38, page: A351, stat: Journal Article,

Endolyn-78, a membrane glycoprotein present in morphologically diverse components of the endosomal and lysosomal compartments: implications for lysosome biogenesis
Croze E; Ivanov IE; Kreibich G; Adesnik M; Sabatini DD; Rosenfeld MG
1989 May;108(5):1597-1613, Journal of cell biology
A monoclonal antibody (2C5) raised against rat liver lysosomal membranes was used to identify a 78-kD glycoprotein that is present in the membranes of both endosomes and lysosomes and, therefore, is designated endolyn-78. In cultures of rat hepatoma (Fu5C8) and kidney cells (NRK), this glycoprotein could not be labeled with [35S]methionine or with [32P]inorganic phosphate but was easily labeled with [35S]cysteine and [3H]mannose. Pulse-chase experiments and determinations of endoglycosidase H (endo H) sensitivity showed that endolyn-78 is derived from a precursor of Mr 58-62 kD that is processed to the mature form with a t1/2 of 15-30 min. The protein has a 22-kD polypeptide backbone that is detected after a brief pulse in tunicamycin-treated cells. During a chase in the presence of the drug, this is converted into an O-glycosylated product of 46 kD that despite the absence of N-linked oligosaccharides is effectively transferred to lysosomes. This demonstrates that the delivery of endolyn-78 to this organelle is not mediated by the mannose-6-phosphate receptor (MPR). Immunocytochemical experiments showed that endolyn-78 is present in the limiting membranes and the interior membranous structures of morphologically identifiable secondary lysosomes that contain the lysosomal hydrolase beta-glucuronidase, lack the MPR, and could not be labeled with alpha-2-macroglobulin at 18.5 degrees C, a temperature which prevents appearance of endocytosed markers in lysosomes. Endolyn-78 was present at low levels in the plasma membrane and in peripheral tubular endosomes, but was prominent in morphologically diverse components of the endosomal compartment (vacuolar endosomes and various types of multivesicular bodies) which acquired alpha-2-macroglobulin at 18.5 degrees C, and frequently contained substantial levels of the MPR and variable levels of beta-glucuronidase. On the other hand, the MPR was very rarely found in endolyn-containing structures that were not labeled with alpha-2-macroglobulin at the low temperature. Thus, the process of lysosomal maturation appears to involve the progressive delivery of lysosomal enzymes to various types of endosomes that may have already received some of the lysosomal membrane proteins. Although endolyn-78 would be one of the proteins added early to endosomes, other lysosomal membrane proteins may be added only to multivesicular endosomes that represent very advanced stages of maturation
— id: 10656, year: 1989, vol: 108, page: 1597, stat: Journal Article,

Characterization of antisera to a synthetic carboxyl-terminal peptide of the glucose transporter protein
Haspel HC; Rosenfeld MG; Rosen OM
1988 Jan 5;263(1):398-403, Journal of biological chemistry
Peptides corresponding to amino acid residues 1-12 of the amino terminal and 480-492 of the carboxyl terminal of the deduced sequence of the glucose transporter were synthesized and used to produce site-specific polyclonal antipeptide sera. In a solid-phase radioimmunoassay, antiserum to the carboxyl terminal recognizes peptide 480-492 and purified human erythrocyte glucose transporter, but not peptide 1-12. Antiserum to the amino terminal recognizes peptide 1-12 but neither peptide 480-492 nor the erythrocyte transporter. The antiserum to the carboxyl terminal specifically immunoblots the Mr 55,000 glucose transporter in erythrocyte membranes and the purified erythrocyte transporter. It also recognizes a Mr 40,000-60,000 polypeptide in membranes of cells derived from different mammalian species and tissues including insulin-sensitive rat adipocytes as well as a Mr 20,000 tryptic fragment of the transporter which contains the site for photolabeling by cytochalasin B. Antiserum to the carboxyl terminal of the transporter binds specifically to leaky erythrocyte membranes but not to intact erythrocytes. This binding is saturable and competitively inhibited by peptide 480-492. Using immunofluorescence microscopy, this antiserum detects glucose transporter protein in permeabilized erythrocytes, but not in intact erythrocytes. These studies provide immunochemical evidence in support of the predicted cytoplasmic orientation of the carboxyl terminus of the glucose transporter, allow us to suggest a spatial relationship of the cytochalasin B binding site to the carboxyl terminal of the glucose transporter and suggest that antisera directed to the carboxyl terminal domain of the protein may be useful for the immunocytochemical localization of the glucose transporter
— id: 65762, year: 1988, vol: 263, page: 398, stat: Journal Article,

CLONING OF A COMPLEMENTARY DNA ENCODING A RAT LIVER POLYPEPTIDE BEARING COMMON ANTIGENIC DETERMINANTS TO A LYSOSOMAL-ENDOSOMAL MEMBRANE PROTEIN
HYMAN C; MORIMOTO T; ADESNIK M; SABATINI D D; ROSENFELD M
1988 ;107(6 PART 3):343A-343A, Journal of cell biology
— id: 104653, year: 1988, vol: 107, page: 343A, stat: Journal Article,

Isolation and characterization of cDNA clones for rat ribophorin I: complete coding sequence and in vitro synthesis and insertion of the encoded product into endoplasmic reticulum membranes
Harnik-Ort V; Prakash K; Marcantonio E; Colman DR; Rosenfeld MG; Adesnik M; Sabatini DD; Kreibich G
1987 Apr;104(4):855-863, Journal of cell biology
Ribophorins I and II are two transmembrane glycoproteins that are characteristic of the rough endoplasmic reticulum and are thought to be part of the apparatus that affects the co-translational translocation of polypeptides synthesized on membrane-bound polysomes. A ribophorin I cDNA clone containing a 0.6-kb insert was isolated from a rat liver lambda gtll cDNA library by immunoscreening with specific antibodies. This cDNA was used to isolate a clone (2.3 kb) from a rat brain lambda gtll cDNA library that contains the entire ribophorin I coding sequence. SP6 RNA transcripts of the insert in this clone directed the in vitro synthesis of a polypeptide of the expected size that was immunoprecipitated with anti-ribophorin I antibodies. When synthesized in the presence of microsomes, this polypeptide, like the translation product of the natural ribophorin I mRNA, underwent membrane insertion, signal cleavage, and co-translational glycosylation. The complete amino acid sequence of the polypeptide encoded in the cDNA insert was derived from the nucleotide sequence and found to contain a segment that corresponds to a partial amino terminal sequence of ribophorin I that was obtained by Edman degradation. This confirmed the identity of the cDNA clone and established that ribophorin I contains 583 amino acids and is synthesized with a cleavable amino terminal insertion signal of 22 residues. Analysis of the amino acid sequence of ribophorin I suggested that the polypeptide has a simple transmembrane disposition with a rather hydrophilic carboxy terminal segment of 150 amino acids exposed on the cytoplasmic face of the membrane, and a luminal domain of 414 amino acids containing three potential N-glycosylation sites. Hybridization measurements using the cloned cDNA as a probe showed that ribophorin I mRNA levels increase fourfold 15 h after partial hepatectomy, in confirmation of measurements made by in vitro translation of liver mRNA. Southern blot analysis of rat genomic DNA suggests that there is a single copy of the ribophorin I gene in the haploid rat genome
— id: 18420, year: 1987, vol: 104, page: 855, stat: Journal Article,

SYNTHESIS AND SORTING TO LYSOSOMES OF BETA-GLUCURONIDASE IN TRANSFECTED CELLS
ANDY, R; ROSENFELD, M; ADESNIK, M; SABATINI, D
1986 NOV ;103(5):A359-A359, Journal of cell biology
— id: 41324, year: 1986, vol: 103, page: A359, stat: Journal Article,

BIOSYNTHESIS OF A CYSTEINE-RICH, HIGHLY GLYCOSYLATED PROTEIN PRESENT IN LYSOSOMAL AND ENDOSOMAL MEMBRANES
CROZE, E; IVANOV, I; SNITKIN, H; KREIBICH, G; SABATINI, DD; ROSENFELD, MG
1986 NOV ;103(5):A355-A355, Journal of cell biology
— id: 41323, year: 1986, vol: 103, page: A355, stat: Journal Article,

PRESENCE OF A SPECIFIC GLYCOPROTEIN IN BOTH ENDOSOMAL AND LYSOSOMAL MEMBRANES
IVANOV, IE; CROZE, E; SNITKIN, H; PLESKEN, H; SABATINI, DD; ROSENFELD, MG
1986 NOV ;103(5):A354-A354, Journal of cell biology
— id: 41322, year: 1986, vol: 103, page: A354, stat: Journal Article,

CHARACTERIZATION AND BIOSYNTHESIS OF A RAT-LIVER MICROSOMAL ESTERASE
MARKS, AS; ROSENFELD, MG; SABATINI, DD; KREIBICH, G
1986 NOV ;103(5):A65-A65, Journal of cell biology
— id: 41318, year: 1986, vol: 103, page: A65, stat: Journal Article,

Nucleotide sequence of rat preputial gland beta-glucuronidase cDNA and in vitro insertion of its encoded polypeptide into microsomal membranes
Nishimura Y; Rosenfeld MG; Kreibich G; Gubler U; Sabatini DD; Adesnik M; Andy R
1986 Oct;83(19):7292-7296, Proceedings of the National Academy of Sciences of the United States of America
We have selected the rat preputial gland beta-glucuronidase as a model protein to study the sorting of newly synthesized lysosomal hydrolases to the lysosome. The complete coding sequence of beta-glucuronidase messenger RNA was determined from the sequences of a group of overlapping cDNA clones isolated from preputial gland cDNA libraries. The beta-glucuronidase mRNA primary translation product contains 648 amino acids, including an amino-terminal signal sequence of 22 residues. The polypeptide has four potential sites for the addition of asparagine-linked core oligosaccharides. A 376-residue segment of beta-glucuronidase shows extensive homology (23% sequence identity) to a portion of Escherichia coli beta-galactosidase. This homology most likely reflects an evolutionary relationship between the bacterial and eukaryotic enzymes and the conservation of structural features necessary for the glycosidase activity of both proteins. Translation of mRNA synthesized in vitro by transcription of a cDNA containing the entire beta-glucuronidase coding region yielded a polypeptide that was immunoprecipitated with anti-beta-glucuronidase antiserum and had the same electrophoretic mobility as the primary translation product of natural beta-glucuronidase mRNA. In the presence of microsomal membranes, the in vitro-synthesized beta-glucuronidase underwent cotranslational incorporation into the microsomes, as indicated by removal of the signal sequence and the addition of several oligosaccharide chains. The beta-glucuronidase cDNA will provide a useful tool to study the mechanism of mannose phosphorylation and other aspects of the sorting of lysosomal enzymes to lysosomes
— id: 18422, year: 1986, vol: 83, page: 7292, stat: Journal Article,

NUCLEOTIDE-SEQUENCE OF RAT PREPUTIAL GLAND BETA-GLUCURONIDASE CDNA AND INVITRO INSERTION OF ITS ENCODED POLYPE
Nishimura, Y; Rosenfeld, MG; Kreibich, G; Gubler, U; Sabatini, DD; Adesnik, M; Andy, R
1986 Dec;11(4):534-534, Cell structure & function
— id: 31421, year: 1986, vol: 11, page: 534, stat: Journal Article,

ISOLATION AND CHARACTERIZATION OF CDNA CLONES FOR RIBOPHORIN-I - COMPLETE CODING SEQUENCE AND INVITRO SYNTHESIS AND INSERTION OF THE ENCODING PRODUCT INTO ER MEMBRANES
ORT, V; PRAKASH, K; COLMAN, DR; ROSENFELD, MG; ADESNIK, M; SABATINI, DD; KREIBICH, G
1986 NOV ;103(5):A65-A65, Journal of cell biology
— id: 41317, year: 1986, vol: 103, page: A65, stat: Journal Article,

Induction of cytochrome P-450 isozymes in rat hepatoma-derived cell cultures
Frey AB; Rosenfeld MG; Dolan WJ; Adesnik M; Kreibich G
1984 Aug;120(2):169-180, Journal of cellular physiology
We have investigated the responsiveness of established rat hepatocyte cell cultures to inducers of cytochrome P-450. One Reuber hepatoma-derived line (Fu5-C8), which under normal culture conditions produces no detectable cytochrome P-450(MC) or cytochrome P-450(PB)--the major cytochrome P-450 isozymes induced by 3-methylcholanthrene and phenobarbital, respectively--was tested for the ability to accumulate either cytochrome P-450 isozyme in response to treatment with various xenobiotics. By immune-precipitation from [35S]-methionine-labeled cell extracts, using monospecific anticytochrome P-450(MC) antibody or monoclonal anticytochrome P-450(PB) antibody, it was demonstrated that these cells possess the capability to synthesize cytochrome P-450(MC) in response to 3-methylcholanthrene treatment, while none of the drug treatments caused the synthesis of detectable quantities of cytochrome P-450(PB). RNA extracted from Fu5-C8 cells directed the in vitro synthesis of immune-precipitable cytochrome P-450(MC) only after treatment of the cells with 3-methylcholanthrene. Kinetic analysis of the response of these cells to 3-methylcholanthrene induction revealed detectable levels of immune-precipitable cytochrome P-450(MC) 2 h after drug treatment with maximal induction occurring between 12 and 16 h of exposure. Another cell line (HF 1.5), obtained originally by hybridization of Fao X H5 variants of a Reuber H35 hepatoma, produces cytochrome P-450(MC) and also cytochrome P-450(PB) constitutively, as determined by specific immune-precipitation from labeled cell extracts. Exposure of confluent monolayers to either phenobarbital or 3-methylcholanthrene resulted in an induction of cytochrome P-450(PB) or cytochrome P-450(MC), respectively. Double-labeling immunofluorescence studies indicate that all cells in the culture produce albumin and most of the cells produce cytochrome P-450(MC), but only a subset of cells synthesize cytochrome P-450(PB). Our results demonstrate that some continuously dividing hepatocyte cell cultures retain the capacity to respond to xenobiotics, including phenobarbital, a response which is typically exhibited by fully differentiated liver cells. Such established hepatocyte cell cultures should prove useful for investigating the mechanism of induction of cytochrome P-450(PB)
— id: 18429, year: 1984, vol: 120, page: 169, stat: Journal Article,

Biosynthesis and processing of ribophorins in the endoplasmic reticulum
Rosenfeld MG; Marcantonio EE; Hakimi J; Ort VM; Atkinson PH; Sabatini D; Kreibich G
1984 Sep;99(3):1076-1082, Journal of cell biology
Ribophorins are two transmembrane glycoproteins characteristic of the rough endoplasmic reticulum, which are thought to be involved in the binding of ribosomes. Their biosynthesis was studied in vivo using lines of cultured rat hepatocytes (clone 9) and pituitary cells (GH 3.1) and in cell-free synthesis experiments. In vitro translation of mRNA extracted from free and bound polysomes of clone 9 cells demonstrated that ribophorins are made exclusively on bound polysomes. The primary translation products of ribophorin messengers obtained from cultured hepatocytes or from regenerating livers co-migrated with the respective mature proteins, but had slightly higher apparent molecular weights (2,000) than the unglycosylated forms immunoprecipitated from cells treated with tunicamycin. This indicates that ribophorins, in contrast to all other endoplasmic reticulum membrane proteins previously studied, contain transient amino-terminal insertion signals which are removed co-translationally. Kinetic and pulse-chase experiments with [35S]methionine and [3H]mannose demonstrated that ribophorins are not subjected to electrophoretically detectable posttranslational modifications, such as proteolytic cleavage or trimming and terminal glycosylation of oligosaccharide side chain(s). Direct analysis of the oligosaccharides of ribophorin l showed that they do not contain the terminal sugars characteristic of complex oligosaccharides and that they range in composition from Man8GlcNAc to Man5GlcNAc. These findings, as well as the observation that the mature proteins are sensitive to endoglycosidase H and insensitive to endoglycosidase D, are consistent with the notion that the biosynthetic pathway of the ribophorins does not require a stage of passage through the Golgi apparatus
— id: 18428, year: 1984, vol: 99, page: 1076, stat: Journal Article,

INCORPORATION OF PROTEINS INTO CELLULAR MEMBRANES AND ORGANELLES
KREIBICH, G; COLMAN, D; ROSENFELD, MG; SABATINI, DD
1983 ;364(9):1164-1165, Hoppe-Seylers zeitschrift fur physiologische Chemie
— id: 40628, year: 1983, vol: 364, page: 1164, stat: Journal Article,

ISOLATION AND CHARACTERIZATION OF CDNA CLONES FOR BETA-GLUCURONIDASE
NISHIMURA, Y; ROSENFELD, M; KREIBICH, G; ADESNIK, M; SABATINI, DD
1983 ;97(5):A103-A103, Journal of cell biology
— id: 40491, year: 1983, vol: 97, page: A103, stat: Journal Article,

Biosynthesis of lysosomal enzymes
Rosenfeld MG; Kreibich G; Sabatini DD; Kato K
1983 ;96(3):764-777, Methods in enzymology
— id: 18432, year: 1983, vol: 96, page: 764, stat: Journal Article,

IDENTIFICATION OF A PROTEIN CHARACTERISTIC OF THE LYSOSOMAL MEMBRANE USING MONOCLONAL-ANTIBODIES
ROSENFELD, M; DOLAN, W; SABATINI, D; KREIBICH, G
1983 ;97(5):A106-A106, Journal of cell biology
— id: 40492, year: 1983, vol: 97, page: A106, stat: Journal Article,

Biosynthesis of lysosomal hydrolases: their synthesis in bound polysomes and the role of co- and post-translational processing in determining their subcellular distribution
Rosenfeld MG; Kreibich G; Popov D; Kato K; Sabatini DD
1982 Apr;93(1):135-143, Journal of cell biology
— id: 18438, year: 1982, vol: 93, page: 135, stat: Journal Article,

Precursor of rat liver alpha 2u-globulin: partial amino acid sequence determination of its signal peptide
Ikehara Y; Oda K; Rosenfeld MG; Bar-Nun S; Kreibich G
1981 Dec;90(6):1833-1836, Journal of biochemistry (Tokyo)
The biosynthesis of alpha 2 u-globulin, the major protein in rat urine, was studied in a cell-free system programmed with messenger RNA isolated from rat liver. Analysis of the primary translation product by SDS-polyacrylamide gel electrophoresis showed a molecular weight of approximatley 22,000 daltons as compared to 20,000 daltons for the mature form. When translation was carried out in the presence of dog pancreas microsomes, alpha 2u-globulin was cotranslationally processed and sequestered in the microsomal lumen. Automated sequencing to mature alpha 2u-globulin and the labeled precursor disclosed the presence of 19 additional amino acids at the amino-terminal end of the primary translation product of the urinary alpha 2u-globulin. The partial amino acid sequence of this extension was determined
— id: 18443, year: 1981, vol: 90, page: 1833, stat: Journal Article,

SYNTHESIS AND COTRANSLATIONAL PROCESSING OF RIBOPHORINS
Rosenfeld, MG; Marcantonio, EE; Harnik, VM; Sabatini, DD; Kreibich, G
1981 ;91(2):A404-A404, Journal of cell biology
— id: 30542, year: 1981, vol: 91, page: A404, stat: Journal Article,

IMMUNOLOGICAL CROSSREACTIVITY OF RAT-LIVER RIBOPHORINS WITH SIMILAR PROTEINS IN OTHER ORGANS AND SPECIES
Marcantonio, EE; Rosenfeld, MG; Sabatini, DD; Kreibich, G
1980 ;87(2):A208-A208, Journal of cell biology
— id: 28088, year: 1980, vol: 87, page: A208, stat: Journal Article,

SYNTHESIS AND MATURATION OF LYSOSOMAL PROTEINS
Rosenfeld, M; Sabatini, DD; Sabban, E; Kato, K; Kreibich, C
1980 ;87(2):A308-A308, Journal of cell biology
— id: 28091, year: 1980, vol: 87, page: A308, stat: Journal Article,