Concomitant preparative isolation of calmodulin and heat shock protein (hsp90) from bovine testes.
We have described a convenient procedure for isolating large amounts of both calmodulin and the 90-kDa heat shock protein (hsp90) from bovine testes. These two proteins coeluted from phenyl-Sepharose during calcium-dependent hydrophobic interaction chromatography. The hsp90 was separated from calmodulin by gel filtration, purified by reverse-phase HPLC or lectin affinity chromatography, and identified by N-terminal amino acid sequence and immunoblotting with hsp90-specific monoclonal antibody. The N-terminus of bovine hsp90 shared 93% amino acid identity with the murine alpha-isoform of hsp90. A third major testes protein of 97 kDa was also isolated with this protocol and was found to be homologous to the 94-kDa glucose-regulated protein.
Wright
L S
LS
Department of Pediatrics, University of Wisconsin Medical School, Madison 53705.
Finn
K A
KA
Siegel
F L
FL
D000595:Amino Acid Sequence; D000818:Animals; D002147:Calmodulin / Q000302:isolation & purification*; D002417:Cattle; D011232:Chemical Precipitation; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006360:Heat-Shock Proteins / Q000302:isolation & purification*; D008297:Male; D008969:Molecular Sequence Data; D013737:Testis / Q000737:chemistry*
D002147:Calmodulin; D006360:Heat-Shock Proteins