Odaka M, Kiribuchi K, Allison W S, Yoshida M
Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan.
FEBS Lett. 1993 Dec 27;336(2):231-5. doi: 10.1016/0014-5793(93)80809-9.
When Tyr-307 of the beta subunit of F1-ATPase from a thermophilic Bacillus strain PS3 is replaced by cysteine and expressed in Escherichia coli cells, about a half population of the mutant beta subunit are labeled by Coenzyme A at Cys-307 through a disulfide bond which is cleavable by reducing treatment. The mutant beta subunit can be reconstituted into the alpha 3 beta 3 complex of which ATPase activity is stimulated two-fold by reducing treatment either prior or after reconstitution. Since Tyr-307 has been supposed to be located at one of subdomains which form the ATP binding site of the beta subunit, Coenzyme A binds to the mutant beta subunit as an AT(D)P analogue in E. coli cells and then covalently attaches to Cys-307.
当嗜热芽孢杆菌菌株PS3的F1 - ATP酶β亚基的酪氨酸- 307被半胱氨酸取代并在大肠杆菌细胞中表达时，约一半的突变β亚基通过可被还原处理裂解的二硫键在半胱氨酸- 307处被辅酶A标记。突变β亚基可重构为α3β3复合体，在重构之前或之后进行还原处理均可使该复合体的ATP酶活性提高两倍。由于酪氨酸- 307被认为位于构成β亚基ATP结合位点的亚结构域之一，辅酶A在大肠杆菌细胞中作为AT（D）P类似物与突变β亚基结合，然后共价连接到半胱氨酸- 307上。
