《MS操作设置_supporting_3》.pdfVIP

Supporting Information for Oct-2-en-4-ynoyl-CoA as a specific inhibitor of acyl-CoA oxidase †,‡ † ‡ Jia Zeng, Long Wu, Xiaojian Zhang, ‡ † *,† Yuandong Liu, Guisheng Deng, and Ding Li † Department of Biology and Chemistry, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong, P. R. China. ‡ Department of Bioengineering, Central South University, Changsha, Hunan 410083, P. R. China *, corresponding author; Tel. (852) 2788 7824; Fax: (852) 2788 7406; E-mail: bhdingli@.hk Materials Acyl-CoA oxidase (ACO) and medium-chain acyl-CoA dehydrogenase (MCAD) were obtained as reported previously (1, 2). Coenzyme A was purchased from Sigma. All other reagents were of research grade or better, and were obtained from commercial sources. Inactivation of ACO by oct-2-en-4-ynoyl-CoA The ACO was protected by 2-octenoyl-CoA (product of ACO catalyzed reaction) from inactivation by oct-2-en-4-ynoyl-CoA as shown in Fig. 1, which indicates the inactivation is active-site directed, and the inhibitor may form a covalent bond with an amino acid residue in the active site of ACO. 100 90 ) 80 % ( 70 y t i v 60 i t c A 50 e v 40 i t a l 30 e R 20 10 0 0 5 10 15 20 Time (min) Fig. 1. Protection by 2-octenoyl-CoA of ACO from inactivation by oct-2-en-4-ynoyl-CoA. The ACO (5