Goat testis calmodulin: purification and physicochemical characterization.
Calmodulin has been purified in large quantities from goat (Capra hiscus) testis. The procedure includes heat treatment, hydrophobic interaction chromatography, and gel filtration. Goat testis calmodulin closely resembles other mammalian testis calmodulin studied so far. The protein has an extinction coefficient value (E1%1cm) of 2.09 at 280 nm, a Stokes radius of 23.2 A at 0.15 M KCl, and a frictional ratio of 1.38. Ca2+, and Tb3+ binding studies demonstrate that the protein has four Ca2(+)-binding sites with a Kd of 52.5 microM. Goat testis calmodulin shows close similarity to other calmodulins in the amino acid composition and in demonstrating an altered migration on SDS/PAGE upon Ca2+ binding. The protein also exhibits anomalously high values for molecular weight and Stokes radius as determined from the analytical gel chromatography and a change in its elution volume with the change of salt concentration in the eluant. These results have been discussed in view of the recently available knowledge from the crystallographic studies of rat testis calmodulin.
Bandyopadhyay
S
S
Crystallography and Molecular Biology Division, Saha Institute of Nuclear Physics, Calcutta, India.
Ghosh
S K
SK
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000737:chemistry / Q000302:isolation & purification*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006041:Goats; D008163:Luminescent Measurements; D008297:Male; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013725:Terbium; D013737:Testis / Q000737:chemistry*
D000596:Amino Acids; D002147:Calmodulin; D013725:Terbium; D002118:Calcium