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Title: Characterization of hyaluronate binding proteins isolated from 3T3 and murine sarcoma virus transformed 3T3 cells

A hyaluronic acid binding fraction was purified from the supernatant media of both 3T3 and murine sarcoma virus (MSV) transformed 3T3 cultures by hyaluronate and immunoaffinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved the hyaluronate affinity-purified fraction into three major protein bands of estimated molecular weight (M/sub r,e/) 70K, 66K, and 56K which contained hyaluronate binding activity and which were termed hyaluronate binding proteins (HABP). Hyaluronate affinity chromatography combined with immunoaffinity chromatography, using antibody directed against the larger HABP, allowed a 20-fold purification of HABP. Fractions isolated from 3T3 supernatant medium also contained additional binding molecules in the molecular weight range of 20K. This material was present in vanishingly small amounts and was not detected with a silver stain or with (/sup 35/S)methionine label. The three protein species isolated by hyaluronate affinity chromatography (M/sub r,e/ 70K, 66K, and 56K) were related to one another since they shared antigenic determinants and exhibited similar pI values. In isocratic conditions, HABP occurred as aggregates of up to 580 kilodaltons. Their glycoprotein nature was indicated by their incorporation of /sup 3/H-sugars. Enzyme-linked immunoadsorbent assay showed they were antigenically distinct from other hyaluronate binding proteins such as fibronectin, cartilage link protein, and the hyaluronate bindingmore » region of chondroitin sulfate proteoglycan. The results are discussed with regard both to the functional significance of hyaluronate-cell surface interactions in transformed as well as normal cells and to the relationship of HABP to other reported hyaluronate binding proteins.« less
Authors:
; ;
Publication Date:
OSTI Identifier:
6127431
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemistry; (United States); Journal Volume: 26:11
Research Org:
Univ. of Calgary, Alberta
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; GLUCOPROTEINS; MOLECULAR STRUCTURE; HYALURONIC ACID; BIOCHEMISTRY; LABELLED COMPOUNDS; BIOCHEMICAL REACTION KINETICS; ELECTROPHORESIS; FIBROBLASTS; METHIONINE; MICE; MOLECULAR WEIGHT; ONCOGENIC VIRUSES; SULFUR 35; AMINES; AMINO ACIDS; ANIMAL CELLS; ANIMALS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CARBOHYDRATES; CARBOXYLIC ACIDS; CHEMISTRY; CONNECTIVE TISSUE CELLS; DAYS LIVING RADIOISOTOPES; DRUGS; EVEN-ODD NUCLEI; ISOTOPES; KINETICS; LIGHT NUCLEI; LIPOTROPIC FACTORS; MAMMALS; MICROORGANISMS; MUCOPOLYSACCHARIDES; NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC SULFUR COMPOUNDS; PARASITES; POLYSACCHARIDES; PROTEINS; RADIOISOTOPES; REACTION KINETICS; RODENTS; SACCHARIDES; SOMATIC CELLS; SULFUR ISOTOPES; VERTEBRATES; VIRUSES 550201* -- Biochemistry-- Tracer Techniques