Lymphocyte function-associated antigen 1 (LFA-1) contains sulfated N-linked oligosaccharides. J Immunol 1985 Jun;134(6):3978-86
Date
06/01/1985Pubmed ID
3886793Scopus ID
2-s2.0-0021804195 (requires institutional sign-in at Scopus site) 26 CitationsAbstract
The murine lymphocyte function-associated antigen 1 (LFA-1) is a glycoprotein heterodimer consisting of an Mr 180,000 alpha-chain and an Mr 95,000 beta-chain. Although LFA-1 has been studied extensively in the past few years due to its involvement in various antigen-specific T lymphocyte responses, virtually nothing is known about its glycosylation. In this report, we have analyzed the oligosaccharide moieties of the murine LFA-1 molecule. Utilizing a T lymphoma cell line, EL-4, it was found that [35S] sulfate, [3H]glucosamine, [3H]mannose, and [3H]fucose were incorporated into both the alpha- and beta-chains of LFA-1. Isolated alpha- and beta-chains from anti-LFA-1 immunoprecipitates of [3H]glucosamine-labeled NP-40 lysates were subjected to tryptic-chymotryptic digestion, and the resulting glycopeptides were fractionated by reverse-phase high performance liquid chromatography. Five major [3H]glucosamine-labeled glycopeptides were generated by this procedure from each of the two polypeptide chains. Treatment of the individual glycopeptides with almond emulsin peptide:N-glycosidase or Endo F demonstrated that the [3H]glucosamine label existed almost entirely in N-linked oligosaccharide structures (Mr 5000 to 10,000). By using similar techniques, the majority of the [35S]sulfate moieties were also found covalently bound to N-linked oligosaccharides. In addition, both [35S]sulfate-labeled alpha- and beta-chains were susceptible to Keratanase and endo-beta-galactosidase digestions, indicating the presence of sulfated N-acetyllactosamine sequences. The expression of [35S]sulfate-labeled LFA-1 on various cell types was also examined. LFA-1 was found to be sulfated only on thymocytes and splenic T cells, but not on macrophages, splenic B, or bone marrow cells.
Author List
Dahms NM, Hart GWAuthor
Nancy M. Dahms PhD Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAntibodies, Monoclonal
Antigens, Surface
Carbohydrate Conformation
Chromatography, High Pressure Liquid
Lymphocyte Function-Associated Antigen-1
Methionine
Mice
Mice, Inbred C3H
Molecular Weight
Oligosaccharides
Rats
Rats, Inbred Strains
Sulfates
Tissue Distribution