Dynamic interaction between components of hexaprenyl diphosphate synthase from Micrococcus luteus BP-26

Abstract

Hexaprenyl diphosphate synthase from Micrococcus luteus BP-26, which has been known to be dissociated into two essential components, designated as components A and B, during hydroxyapatite chromatography [Fujii, H., Koyama, T., & Ogura, K. (1982) J. Biol. Chem. 257, 14610-14612], was also resolved similarly by Sephadex G-100 or DEAE ion-exchange chromatography. Each component takes various self-aggregated forms. The apparent molecular mass of component B estimated by gel filtration on Superose 12 varied depending on its concentration, ranging from approximately 18 to 49 kilodaltons (kDa). On the other hand, the apparent molecular mass of component A varied depending on not only its concentration but also the ionic strength of the medium, ranging from approximately 13 to 24 kDa. When a mixture of components A and B preincubated in the presence of Mg2+ but in the absence of substrate was subjected to Superose 12 gel filtration, they were eluted at positions identical with those observed when they were chromatographed individually. In contrast, when a mixture of components A and B incubated in the presence of Mg2+ and substrates was filtrated on Superose 12, the elution positions were markedly changed, showing that an approximately 24-kDa aggregate of component A (designated as A) and an approximately 27-kDa aggregate of component B (designated as B) were the major species. Evidence was also obtained to show that farnesyl diphosphate (FPP) binds to the components to form an aggregate, A-B-FPP-Mg2+, which probably represents an intermediary state of enzyme catalysis.