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.