Identification and characterization of heparan sulfate-binding proteins from human lung carcinoma cells.
The heparan sulfate proteoglycan/heparin-binding proteins of the human lung carcinoma cell line LX-1 have been identified, partially purified, and characterized. Analysis of the binding of [3H]heparin to membranes isolated from LX-1 cells indicated the presence of two classes of binding sites, with Kd values of approximately 2 x 10(-10) and 4 x 10(-8) M and corresponding Bmax values of 1 x 10(5) and 2 x 10(7) binding sites/cell. Binding was also observed with isolated heparan sulfate chains and with intact heparan sulfate proteoglycan isolated from two different cell types. With each ligand, binding was inhibited by addition of unlabeled heparin. The binding proteins were extracted from LX-1 cell membranes in detergent solution, and two size classes of binding proteins were identified by overlaying transblots of electrophoretically separated proteins with radioactive ligands. These two classes of binding proteins were shown to contain doublets with estimated molecular masses of approximately 16 kDa (HSBP1A and HSBP1B) and approximately 32 kDa (HSBP2A and HSBP2B). The proteins were partially purified by heparin-Sepharose chromatography and shown to bind heparin and heparan sulfate proteoglycan. By amino acid composition, N-terminal amino acid sequence, and reactivity with antibody, HSBP1A was shown to be very similar to histone 2B; HSBP1B may also be related to histone 2A. HSBP2A and HSBP2B, however, did not react with antibodies to the major histones and had compositions different from one another and from HSBP1.
Bilozur
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Department of Anatomy and Cellular Biology, Tufts University Health Science Schools, Boston, Massachusetts 02111.
Biswas
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D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis* / Q000378:metabolism; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D019812:Heparan Sulfate Proteoglycans; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D002352:Carrier Proteins; D011508:Chondroitin Sulfate Proteoglycans; D003101:Collodion; D019812:Heparan Sulfate Proteoglycans; D006493:Heparin; D006497:Heparan Sulfate