Molecular weight and charge heterogeneity of Thy-1 glycoprotein in different populations of T-cells

M. H. Morrison, W. G. Chaney, W. J. Esselman

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Molecular weight and charge microheterogeneity of Thy-1 glycoprotein expressed on different T-cell populations were compared by two-dimensional gel electrophoresis and endoglycosidase treatment. Thy-1 was immunoprecipitated from detergent lysates of radioiodinated T-cells of spleens, lymph nodes, Peyer's patches, peripheral blood, IL-2-cultured thymocytes and peanut agglutinin (PNA) separated thymocytes. In general, thymocytes and IL-2-cultured thymocytes expressed the highest levels of Thy-1 with a large Mr and charge variation. The Thy-1 of these cells was basic and contained low levels of sialic acid. On the other hand, peripheral T-lymphocytes exhibited a restricted Mr and more limited charge heterogeneity with higher amounts of sialic acid. The Thy-1 glycoprotein of mature, low Thy-1 PNA- thymocytes had Mr and charge heterogeneity identical to peripheral T-lymphocytes. The Mr and charge variation of immature PNA+ thymocytes was essentially identical to that of whole thymocytes. The molecular source of Mr heterogeneity in thymocyte Thy-1 and restricted Mr, in lymph node Thy-1 was studied by analysis with endoglycosidase-H (Endo-H) and Endo-D. The results of Endo-H digestion showed that most of the Thy-1 polypeptides from both thymocyte and lymph node contained one high-mannose type oligosaceharide which was relatively small and not heterogeneous with respect to Mr. The complex-type oligosaccharides (Endo-D-sensitive) were larger and were responsible for the broad Mr-heterogeneity found in thymocyte Thy-1. Both thymocyte and lymph node Thy-1 generally appear to contan three N-linked oligosaccharides (one high-mannose and two complex) and sequential hydrolysis with Endo-H and Endo-D resulted in an unglycosylated polypeptide with an Mr of approx. 12,500.

Original languageEnglish (US)
Pages (from-to)405-413
Number of pages9
JournalMolecular Immunology
Volume21
Issue number5
DOIs
StatePublished - May 1984

Fingerprint

Thymocytes
Glycoproteins
Molecular Weight
T-Lymphocytes
Population
Glycoside Hydrolases
Peanut Agglutinin
Lymph Nodes
N-Acetylneuraminic Acid
Mannose
Oligosaccharides
Interleukin-2
Peyer's Patches
Peptides
Electrophoresis, Gel, Two-Dimensional
Detergents
Digestion
Hydrolysis
Spleen

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

Cite this

Molecular weight and charge heterogeneity of Thy-1 glycoprotein in different populations of T-cells. / Morrison, M. H.; Chaney, W. G.; Esselman, W. J.

In: Molecular Immunology, Vol. 21, No. 5, 05.1984, p. 405-413.

Research output: Contribution to journalArticle

Morrison, M. H. ; Chaney, W. G. ; Esselman, W. J. / Molecular weight and charge heterogeneity of Thy-1 glycoprotein in different populations of T-cells. In: Molecular Immunology. 1984 ; Vol. 21, No. 5. pp. 405-413.
@article{cedfdd6fbd714532b1a3c6f46c85fefc,
title = "Molecular weight and charge heterogeneity of Thy-1 glycoprotein in different populations of T-cells",
abstract = "Molecular weight and charge microheterogeneity of Thy-1 glycoprotein expressed on different T-cell populations were compared by two-dimensional gel electrophoresis and endoglycosidase treatment. Thy-1 was immunoprecipitated from detergent lysates of radioiodinated T-cells of spleens, lymph nodes, Peyer's patches, peripheral blood, IL-2-cultured thymocytes and peanut agglutinin (PNA) separated thymocytes. In general, thymocytes and IL-2-cultured thymocytes expressed the highest levels of Thy-1 with a large Mr and charge variation. The Thy-1 of these cells was basic and contained low levels of sialic acid. On the other hand, peripheral T-lymphocytes exhibited a restricted Mr and more limited charge heterogeneity with higher amounts of sialic acid. The Thy-1 glycoprotein of mature, low Thy-1 PNA- thymocytes had Mr and charge heterogeneity identical to peripheral T-lymphocytes. The Mr and charge variation of immature PNA+ thymocytes was essentially identical to that of whole thymocytes. The molecular source of Mr heterogeneity in thymocyte Thy-1 and restricted Mr, in lymph node Thy-1 was studied by analysis with endoglycosidase-H (Endo-H) and Endo-D. The results of Endo-H digestion showed that most of the Thy-1 polypeptides from both thymocyte and lymph node contained one high-mannose type oligosaceharide which was relatively small and not heterogeneous with respect to Mr. The complex-type oligosaccharides (Endo-D-sensitive) were larger and were responsible for the broad Mr-heterogeneity found in thymocyte Thy-1. Both thymocyte and lymph node Thy-1 generally appear to contan three N-linked oligosaccharides (one high-mannose and two complex) and sequential hydrolysis with Endo-H and Endo-D resulted in an unglycosylated polypeptide with an Mr of approx. 12,500.",
author = "Morrison, {M. H.} and Chaney, {W. G.} and Esselman, {W. J.}",
year = "1984",
month = "5",
doi = "10.1016/0161-5890(84)90038-5",
language = "English (US)",
volume = "21",
pages = "405--413",
journal = "Molecular Immunology",
issn = "0161-5890",
publisher = "Elsevier Limited",
number = "5",

}

TY - JOUR

T1 - Molecular weight and charge heterogeneity of Thy-1 glycoprotein in different populations of T-cells

AU - Morrison, M. H.

AU - Chaney, W. G.

AU - Esselman, W. J.

PY - 1984/5

Y1 - 1984/5

N2 - Molecular weight and charge microheterogeneity of Thy-1 glycoprotein expressed on different T-cell populations were compared by two-dimensional gel electrophoresis and endoglycosidase treatment. Thy-1 was immunoprecipitated from detergent lysates of radioiodinated T-cells of spleens, lymph nodes, Peyer's patches, peripheral blood, IL-2-cultured thymocytes and peanut agglutinin (PNA) separated thymocytes. In general, thymocytes and IL-2-cultured thymocytes expressed the highest levels of Thy-1 with a large Mr and charge variation. The Thy-1 of these cells was basic and contained low levels of sialic acid. On the other hand, peripheral T-lymphocytes exhibited a restricted Mr and more limited charge heterogeneity with higher amounts of sialic acid. The Thy-1 glycoprotein of mature, low Thy-1 PNA- thymocytes had Mr and charge heterogeneity identical to peripheral T-lymphocytes. The Mr and charge variation of immature PNA+ thymocytes was essentially identical to that of whole thymocytes. The molecular source of Mr heterogeneity in thymocyte Thy-1 and restricted Mr, in lymph node Thy-1 was studied by analysis with endoglycosidase-H (Endo-H) and Endo-D. The results of Endo-H digestion showed that most of the Thy-1 polypeptides from both thymocyte and lymph node contained one high-mannose type oligosaceharide which was relatively small and not heterogeneous with respect to Mr. The complex-type oligosaccharides (Endo-D-sensitive) were larger and were responsible for the broad Mr-heterogeneity found in thymocyte Thy-1. Both thymocyte and lymph node Thy-1 generally appear to contan three N-linked oligosaccharides (one high-mannose and two complex) and sequential hydrolysis with Endo-H and Endo-D resulted in an unglycosylated polypeptide with an Mr of approx. 12,500.

AB - Molecular weight and charge microheterogeneity of Thy-1 glycoprotein expressed on different T-cell populations were compared by two-dimensional gel electrophoresis and endoglycosidase treatment. Thy-1 was immunoprecipitated from detergent lysates of radioiodinated T-cells of spleens, lymph nodes, Peyer's patches, peripheral blood, IL-2-cultured thymocytes and peanut agglutinin (PNA) separated thymocytes. In general, thymocytes and IL-2-cultured thymocytes expressed the highest levels of Thy-1 with a large Mr and charge variation. The Thy-1 of these cells was basic and contained low levels of sialic acid. On the other hand, peripheral T-lymphocytes exhibited a restricted Mr and more limited charge heterogeneity with higher amounts of sialic acid. The Thy-1 glycoprotein of mature, low Thy-1 PNA- thymocytes had Mr and charge heterogeneity identical to peripheral T-lymphocytes. The Mr and charge variation of immature PNA+ thymocytes was essentially identical to that of whole thymocytes. The molecular source of Mr heterogeneity in thymocyte Thy-1 and restricted Mr, in lymph node Thy-1 was studied by analysis with endoglycosidase-H (Endo-H) and Endo-D. The results of Endo-H digestion showed that most of the Thy-1 polypeptides from both thymocyte and lymph node contained one high-mannose type oligosaceharide which was relatively small and not heterogeneous with respect to Mr. The complex-type oligosaccharides (Endo-D-sensitive) were larger and were responsible for the broad Mr-heterogeneity found in thymocyte Thy-1. Both thymocyte and lymph node Thy-1 generally appear to contan three N-linked oligosaccharides (one high-mannose and two complex) and sequential hydrolysis with Endo-H and Endo-D resulted in an unglycosylated polypeptide with an Mr of approx. 12,500.

UR - http://www.scopus.com/inward/record.url?scp=0021215248&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021215248&partnerID=8YFLogxK

U2 - 10.1016/0161-5890(84)90038-5

DO - 10.1016/0161-5890(84)90038-5

M3 - Article

C2 - 6146094

AN - SCOPUS:0021215248

VL - 21

SP - 405

EP - 413

JO - Molecular Immunology

JF - Molecular Immunology

SN - 0161-5890

IS - 5

ER -