Nucleoside Triphosphate(NTP)-Binding Proteins and Endogenous ADP-ribosyl Transferase in Neurospora crassa
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Nucleoside triphosphate(NTP)-binding proteins were detected in the crude extract of mycelia of Neurospora crassa, which was treated with 1% Lubrol PX and fractionated by gel filtration. Protein fractions showing the capacity to bind [^<35>S]ATPγS or [^<35>S]GTPγS were designated as ANG1 to 6. The binding of [^<35>S]ATPγS or [^<35>S]GTPγS was prevented in the presence of 0.1 mM ATP or GTP except that in fractions ANG1 and 2, the presence of GTP stimulated the binding of [^<35>S]ATPγS to ATP(NTP)-binding proteins. ATP or GTP was 1 to 2 orders of magnitude more effective than CTP or UTP in preventing the binding of [^<35>S]GTPγS in ANG1, 2 and 5. Among these fractions ANG1, 2, 5 and 6 showed activity to hydrolyze 1 nM [γ-^<32>P]ATP or [γ-^<32>P]GTP. NTP-binding proteins bound with [^<35>S]ATPγS or [^<35>S]GTPγS had lower apparent molecular weights than the same proteins without bound nucleotide. Proteins bound with [^<35>S]ATPγS or [^<35>S]GTPγS and those [^<32>P]ADP-ribosylated by endogenous ADP-ribosyl transferase in each fraction were analyzed by SDS-PAGE. About 20 species of ATP or ATP-GTP-binding proteins were detected, several of which were ADP-ribosylated. The binding of [^<35>S]ATPγS or [^<35>S]-GTPγS to NTP-binding proteins was confirmed by the comparison of non-boiled and boiled samples immediately before loading to SDS-PAGE. ATP, GTP, CTP or UTP at the concentration of O.1 mM effectively removed [^<35>S]ATPγS or [^<35>S]GTPγS bound to NTP-binding proteins.
- 日本植物生理学会の論文
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