Growth stimulating activity of heat shock protein 90 alpha to lymphoid cell lines in serum-free medium.
A growth stimulating factor was purified from the culture supernatant of human-human hybridoma SH-76 cells in serum-free RPMI 1640 medium by the serial use of DEAE anion and heparin affinity chromatographies. The factor was a 85 kDa protein on SDS-polyacrylamide gel electrophoresis. N-terminal amino acid sequence (PEETQTQDQPME) of the protein was coincident with that of heat shock protein 90 alpha (HSP90 alpha). The protein reacted with anti-HSP90 monoclonal antibody. These results suggest that the protein was a member of HSP90 alpha family after taking other circumstantial evidence into account. The protein stimulated the growth of some lymphoid cell lines such as human B-lymphoblastoid cell line HO-323-3 hybridomas derived from HO-323, and several other lymphoid cell lines. There were several lymphoid cell lines which did not respond to the protein. Growth stimulating activity of the protein was heat-unstable and significantly decreased at above 60 degrees C. These are the first data that describe growth-stimulating activity of heat shock protein 90 alpha.
Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, Japan.
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T
Kuroita
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Tachibana
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H
Ohashi
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S
Shirahata
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H
Murakami
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002455:Cell Division; D002460:Cell Line; D017077:Culture Media, Conditioned / Q000737:chemistry; D016895:Culture Media, Serum-Free; D006133:Growth Substances / Q000302:isolation & purification / Q000502:physiology*; D006360:Heat-Shock Proteins / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D006825:Hybridomas; D008214:Lymphocytes / Q000166:cytology*; D051379:Mice; D008969:Molecular Sequence Data
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D006133:Growth Substances; D006360:Heat-Shock Proteins