Purification of homogeneous gamma-cystathionase and study of its structure by circular dichroism
Lupu, E.I.; Bolotina, I.A.; Goriachenkova, E.V.
Molekuliarnaia Biologiia 19(1): 67-74
1985
ISSN/ISBN: 0026-8984 PMID: 3982410 Document Number: 263276
Rat liver gamma-cystathionase has been purified to homogeneity (verified by SDS electrophoresis and ultracentrifugation). The secondary and tertiary structures of the enzyme were studied by circular dichroism spectra. Our studies revealed that the holoenzyme molecule comprises approximately 22% of alpha-helices, 14% of beta-structure, 14% of beta-bends, and 50% of unordered structure. Conformational alterations of the enzyme molecule resulting from enzyme PLP elimination, reduction with sodium borohydride and irreversible inhibition by propargylglycine were examined. The enzyme's secondary structure was shown to be stable whereas the tertiary structure is labile. Saturation with PLP maintains the enzyme's optimal (catalytically active) tridimensional structure. Sodium dodecylsulfate alters its secondary (the amount of alpha-helix being raised to 34%) and tertiary structures.