Carbohydrate moeity of HLA antigens. Antigenic properties and amino acid sequences around the site of glycosylation

P. Parham, B. N. Alpert, Harry T Orr, J. L. Strominger

Research output: Contribution to journalArticle

102 Citations (Scopus)

Abstract

The carbohydrate moiety of HLA antigens was radioactively labeled in galactose residues by treatment of membranes with neuraminidase, galactose oxidase, and NaB3H4. A single N-linked oligosaccharide side chain M(r) 2700 to 3300 was labeled by this treatment. Unique pronase and tryptic 3H-glycopeptides were purified, using lentil-lectin affinity chromatography. They contained all the radioactivity and all the carbohydrate of the HLA heavy chain. The properties on gel filtration and high voltage electrophoresis of the pronase 3H-glycopeptides from HLA-A2 and HLA-B7 were identical, whereas the tryptic 3H-glycopeptides showed significant differences in both size and charge. The tryptic glycopeptide preparation had less than 0.1% of the HLA antigenic activity of the intact molecule. Removal by glycosidases of all the carbohydrate, except for single residues of N-acetylglucosamine and fucose, from the intact molecule was accompanied by no change in HLA antigenic activity. These data show that the carbohydrate plays no direct role in determining the antigenic specificity of HLA antigens. The single site of glycosylation in the HLA heavy chain was localized to a position about 100 residues from the NH2-terminal end. Amino acid sequences of tryptic 3H-glycopeptides from HLA-A2 and HLA-B7 were highly homologous. These sequences also showed homology with variable regions of human immunoglobulin heavy chains and with NH2-terminal sequences of HLA heavy chains. This suggests that HLA heavy chains are structurally related to immunoglobulins and made up of structural homology units.

Original languageEnglish (US)
Pages (from-to)7555-7567
Number of pages13
JournalJournal of Biological Chemistry
Volume252
Issue number21
StatePublished - Jan 1 1977
Externally publishedYes

Fingerprint

Glycosylation
Glycopeptides
HLA Antigens
Amino Acid Sequence
Carbohydrates
Amino Acids
HLA-B7 Antigen
HLA-A2 Antigen
Pronase
Galactose Oxidase
Affinity chromatography
Immunoglobulin Heavy Chains
Molecules
Fucose
Acetylglucosamine
Glycoside Hydrolases
Radioactivity
Neuraminidase
Electrophoresis
Oligosaccharides

Cite this

Carbohydrate moeity of HLA antigens. Antigenic properties and amino acid sequences around the site of glycosylation. / Parham, P.; Alpert, B. N.; Orr, Harry T; Strominger, J. L.

In: Journal of Biological Chemistry, Vol. 252, No. 21, 01.01.1977, p. 7555-7567.

Research output: Contribution to journalArticle

@article{0f4e7e1c378c4911be1bc3f70c4b5e23,
title = "Carbohydrate moeity of HLA antigens. Antigenic properties and amino acid sequences around the site of glycosylation",
abstract = "The carbohydrate moiety of HLA antigens was radioactively labeled in galactose residues by treatment of membranes with neuraminidase, galactose oxidase, and NaB3H4. A single N-linked oligosaccharide side chain M(r) 2700 to 3300 was labeled by this treatment. Unique pronase and tryptic 3H-glycopeptides were purified, using lentil-lectin affinity chromatography. They contained all the radioactivity and all the carbohydrate of the HLA heavy chain. The properties on gel filtration and high voltage electrophoresis of the pronase 3H-glycopeptides from HLA-A2 and HLA-B7 were identical, whereas the tryptic 3H-glycopeptides showed significant differences in both size and charge. The tryptic glycopeptide preparation had less than 0.1{\%} of the HLA antigenic activity of the intact molecule. Removal by glycosidases of all the carbohydrate, except for single residues of N-acetylglucosamine and fucose, from the intact molecule was accompanied by no change in HLA antigenic activity. These data show that the carbohydrate plays no direct role in determining the antigenic specificity of HLA antigens. The single site of glycosylation in the HLA heavy chain was localized to a position about 100 residues from the NH2-terminal end. Amino acid sequences of tryptic 3H-glycopeptides from HLA-A2 and HLA-B7 were highly homologous. These sequences also showed homology with variable regions of human immunoglobulin heavy chains and with NH2-terminal sequences of HLA heavy chains. This suggests that HLA heavy chains are structurally related to immunoglobulins and made up of structural homology units.",
author = "P. Parham and Alpert, {B. N.} and Orr, {Harry T} and Strominger, {J. L.}",
year = "1977",
month = "1",
day = "1",
language = "English (US)",
volume = "252",
pages = "7555--7567",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "21",

}

TY - JOUR

T1 - Carbohydrate moeity of HLA antigens. Antigenic properties and amino acid sequences around the site of glycosylation

AU - Parham, P.

AU - Alpert, B. N.

AU - Orr, Harry T

AU - Strominger, J. L.

PY - 1977/1/1

Y1 - 1977/1/1

N2 - The carbohydrate moiety of HLA antigens was radioactively labeled in galactose residues by treatment of membranes with neuraminidase, galactose oxidase, and NaB3H4. A single N-linked oligosaccharide side chain M(r) 2700 to 3300 was labeled by this treatment. Unique pronase and tryptic 3H-glycopeptides were purified, using lentil-lectin affinity chromatography. They contained all the radioactivity and all the carbohydrate of the HLA heavy chain. The properties on gel filtration and high voltage electrophoresis of the pronase 3H-glycopeptides from HLA-A2 and HLA-B7 were identical, whereas the tryptic 3H-glycopeptides showed significant differences in both size and charge. The tryptic glycopeptide preparation had less than 0.1% of the HLA antigenic activity of the intact molecule. Removal by glycosidases of all the carbohydrate, except for single residues of N-acetylglucosamine and fucose, from the intact molecule was accompanied by no change in HLA antigenic activity. These data show that the carbohydrate plays no direct role in determining the antigenic specificity of HLA antigens. The single site of glycosylation in the HLA heavy chain was localized to a position about 100 residues from the NH2-terminal end. Amino acid sequences of tryptic 3H-glycopeptides from HLA-A2 and HLA-B7 were highly homologous. These sequences also showed homology with variable regions of human immunoglobulin heavy chains and with NH2-terminal sequences of HLA heavy chains. This suggests that HLA heavy chains are structurally related to immunoglobulins and made up of structural homology units.

AB - The carbohydrate moiety of HLA antigens was radioactively labeled in galactose residues by treatment of membranes with neuraminidase, galactose oxidase, and NaB3H4. A single N-linked oligosaccharide side chain M(r) 2700 to 3300 was labeled by this treatment. Unique pronase and tryptic 3H-glycopeptides were purified, using lentil-lectin affinity chromatography. They contained all the radioactivity and all the carbohydrate of the HLA heavy chain. The properties on gel filtration and high voltage electrophoresis of the pronase 3H-glycopeptides from HLA-A2 and HLA-B7 were identical, whereas the tryptic 3H-glycopeptides showed significant differences in both size and charge. The tryptic glycopeptide preparation had less than 0.1% of the HLA antigenic activity of the intact molecule. Removal by glycosidases of all the carbohydrate, except for single residues of N-acetylglucosamine and fucose, from the intact molecule was accompanied by no change in HLA antigenic activity. These data show that the carbohydrate plays no direct role in determining the antigenic specificity of HLA antigens. The single site of glycosylation in the HLA heavy chain was localized to a position about 100 residues from the NH2-terminal end. Amino acid sequences of tryptic 3H-glycopeptides from HLA-A2 and HLA-B7 were highly homologous. These sequences also showed homology with variable regions of human immunoglobulin heavy chains and with NH2-terminal sequences of HLA heavy chains. This suggests that HLA heavy chains are structurally related to immunoglobulins and made up of structural homology units.

UR - http://www.scopus.com/inward/record.url?scp=0017622587&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017622587&partnerID=8YFLogxK

M3 - Article

C2 - 72068

AN - SCOPUS:0017622587

VL - 252

SP - 7555

EP - 7567

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 21

ER -