Purification and partial characterization of two glycoproteins in bovine peripheral nerve myelin membrane

Kïtamura, Kazuo; Suzuki, Masaru; Uyemura, Keiichi

doi:10.1016/0005-2736(76)90050-x

Cited by 157 publications

(86 citation statements)

References 27 publications

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“…2C); the few exceptions may represent species differences between bovine PAS-II and rat SR13. PAS-II was isolated from purified myelin of various species (23) and shows the same developmental pattern of expression in the chicken sciatic nerve as other major myelin proteins (25). Based on these results, we conclude that PAS-II and SR13 are likely to be the same myelin protein.…”

Section: Discussionmentioning

confidence: 53%

A myelin protein is encoded by the homologue of a growth arrest-specific gene.

Welcher

Suter

León

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

199

143

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Striking features of the cellular response to sciatic nerve injury are the proliferation of Schwann cells in the distal nerve stump and the downregulation of myelin-specific gene expression. Once the axons regrow, the Schwann cells differentiate again to reform the myelin sheaths. We have isolated a rat cDNA, SR13, which is strongly downregulated in the initial phase after sciatic nerve injury. This cDNA encodes a glycoprotein that shares striking amino acid similarity with a purified myelin protein and is specifically precipitated by a myelin-specific antiserum. Immunohistochemistry experiments using peptide-specific polyclonal antibodies localize the SR13 protein to the myelin sheath of the sciatic nerve. Computer-aided sequence analysis identified a pronounced homology of SR13 to a growth arrest-specific mRNA (Gas-3) that is expressed in resting but not in proliferating 3T3 mouse fibroblasts. SR13 is similarly downregulated during Schwann cell proliferation in the rat sciatic nerve. The association of the SR13 as well as the Gas-3 mRNA with nonproliferating cells in two different experimental systems suggests a common role for these molecules in maintaining the quiescent cell state.

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Section: Discussionmentioning

confidence: 53%

A myelin protein is encoded by the homologue of a growth arrest-specific gene.

Welcher

Suter

León

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

199

143

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“…(9) Po were determined by electrophoresis on 12.5% polyacrylamide gels made by the method of Laemmli (10). The P2 and Po antisera were prepared in rabbits by multiple injections of 1-mg quantities of each protein in complete Freund's adjuvant.…”

Section: Methodsmentioning

confidence: 99%

Immunocytochemical localization of rat peripheral nervous system myelin proteins: P2 protein is not a component of all peripheral nervous system myelin sheaths.

Trapp¹,

McIntyre²,

Quarles³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

114

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Specific antibodies have been developed against PI, P2, and Po myelin proteins and were used to study the localization of these proteins in the rat peripheral nervous system. Both peripheral and central nervous system myelin sheaths contain Pi protein. Po and P2 proteins are found exclusively in peripheral nervous system myelin sheaths. Antisera to Pi and Po proteins stain all peripheral nervous system myelin sheaths uniformly. P2 protein is not a component of all peripheral nervous system myelin sheaths. In sheaths that do contain P2 protein, it is concentrated in the area of the Schmidt-Lanterman incisures.Myelin isolated from mammalian peripheral nervous system (PNS) contains three major proteins (1). These proteins include a glycoprotein, Po, with a molecular weight (Mr) of 30,000 and two basic proteins, PI and P2, with Mr of 18,500 and 13,500, respectively. The Po and P2 proteins are unique to PNS myelin whereas the P1 protein appears to be identical to the large central nervous system (CNS) myelin basic protein (2). The Po protein represents approximately 50% of total PNS myelin protein whereas the quantities of PI and P2 and the ratio of P1/P2 varies from species to species (1). The ratio of PI/P2 also varies within different PNS fiber tracts obtained from the same animals (1). Myelin purification provides a heterogeneous population of myelin membranes. By virtue of the myelin isolation procedure, anatomical specificity of myelin proteins within individual myelinated fibers or myelin internodes is lost prior to biochemical analysis. Indirect evidence has localized the Po protein at the intraperiod line (3-5) and the basic proteins at the major dense line (5-7) of myelin. These studies have not defined the anatomical distribution of the major PNS myelin proteins within myelin sheaths of PNS fiber tracts.In the present study we present light microscopic immunocytochemical evidence that in the rat P2 protein is not a component of all PNS-myelin sheaths, but in fact is a component of only some myelin sheaths. In addition, when antiserum to P2 stains a myelin sheath, Schmidt-Lanterman clefts stain more intensely than compact portions of the internode. Po and PI proteins are located in all PNS myelin sheaths thus far analyzed, and antisera to them stain all sheaths uniformly. MATERIALS AND METHODSSeven-day-old, 25-day-old, and adult (200-225 g) SpragueDawley rats were anesthetized with ether and fixed by intracardiac perfusion with a solution containing 76 ml of saturated HgCl2 and 20 ml of 37% (vol/vol) formaldehyde. Trigeminal

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“…data] anti-PO reactive band. The P0 protein has a single glycosylation site [34], and the carbohydrate portion of the P0 protein is, like the G1N1 epitope, N-linked; it has 3 residues each of mannose and N-acetylglucosamine, and 1 residue each of galac tose, sialic acid and fucose [34,35]. The P0 protein has also been shown by others [36] to express the L2/HNK-1 carbohydrate epi tope.…”

Section: Discussionmentioning

confidence: 99%

Expression of GlN1-Immunoreactive Glycoproteins during Development and Regeneration in Avian Peripheral Nerve

Snyder

Barbu²,

Uzman³

1990

Dev Neurosci

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Reactivity of the monoclonal antibody GlN1 was examined by immunocytochemistry and immunoblotting in quail and chick developing and regenerating wing nerves; its distribution is compared to that of HNK-1 which detects a carbohydrate epitope widely distributed in the nervous system. Reactivity was detected by immunofluorescence in cryostat sections, by a postembedding electron-microscopic immunogold technique and in immunoblots of nerve homogenates. From E11-E16, reactivity was detected in several large (120–260 kD) glycoprotein bands; and thereafter, principally, in 3 myelin-related glycoproteins (100, 26.5, and 21.5/19.5 kD). The HNK-1 carbohydrate epitope was detected in all these and in other bands permitting identification of the 100-kD moiety as the myelin-associated glycoprotein and the 26.5-kD protein as the P0 protein; the myelin-related 21.5/19.5-kD doublet appears as distinct, probable adhesion molecule(s). During development and regeneration, GlN1 reactivity detected by immunogold labeling of sections appeared first over the extracellular matrix, and later over thicker myelin sheaths. After transection, immunoreactivity in immunoblots was lost in distal stumps but reappeared with time; first in the larger (> 120 kD) moieties and then in the myelin-related bands, the same sequence observed in development. Electron-microscopic detection of both GlN1 and HNK-1 carbohydrate epitopes by immunogold labeling of resin-embedded sections is localized most consistently in the thicker (>0.3 µm) myelin sheaths of nerves from chicks at or after hatching. Immunoblots of mature fowl nerve tissues homogenized at various stages of preparation for electron microscopy (after fixation or after fixation and dehydration) show sustained immunoreactivity in the 21.5/19.5-kD bands, reduction or complete suppression in others, and evidence of immunoreactivity in high molecular weight, presumably cross-linked, constituents that remain in the stacking gel portion of the blots.

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Purification and partial characterization of two glycoproteins in bovine peripheral nerve myelin membrane

Cited by 157 publications

References 27 publications

A myelin protein is encoded by the homologue of a growth arrest-specific gene.

A myelin protein is encoded by the homologue of a growth arrest-specific gene.

Immunocytochemical localization of rat peripheral nervous system myelin proteins: P2 protein is not a component of all peripheral nervous system myelin sheaths.

Expression of GlN1-Immunoreactive Glycoproteins during Development and Regeneration in Avian Peripheral Nerve

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