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.
N1 - Funding Information:
Knowledge of surface antigens and receptors facilitates the study of cellular differentiation and functional maturation of lymphocytes. Increased residues of sialic acid and oligosaccharides, as well as the modulation of relative levels of antigens such as H-2, Thy-l, TL and Lyt-1,2,3, accompany maturation of T-cells in the thymus and migration of T-ceils to peripheral lymphoid organs (Reisner et al., 1980; Williams, 1982; Hoessli et al., 1980~). Since Thy-l is an abundant, membranous glycoprotein found on thymocytes, and since its density on T-cells decreases during maturation, it is of interest to compare the carbohydrate differences between thymocyte and peripheral lymphocyte Thy-1. Thy-l was originally described as a differentiation antigen of thymus-derived lymphocytes and brain cells (Reif and Allen, 1966). Recent studies have shown that the antigen is also present on bone marrow hematopoietic cells (Basch and Berman, *This investigation was supported by a PHS Grant, No. CA24437, awarded by the National Cancer Institute, DHHS. fAddress correspondence to this author. ~Abbrev~ations: PNA, peanut agglutinin; PBS, phosphate-buffered saline; SDS-gel, sodium dodecylsulfate-_ nolyacrvlamide . gel; NEPHGE, non-equilibrium pH-gel
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.
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U2 - 10.1016/0161-5890(84)90038-5
DO - 10.1016/0161-5890(84)90038-5
M3 - Article
C2 - 6146094
AN - SCOPUS:0021215248
SN - 0161-5890
VL - 21
SP - 405
EP - 413
JO - Molecular Immunology
JF - Molecular Immunology
IS - 5
ER -