Binding of camphor to Pseudomonas putida cytochrome P450cam: steady‐state and picosecond time‐resolved fluorescence studies

FEBS Letters

Mitra, Samaresh¹  Prasad, Swati¹  Mazumdar, Shyamalava¹ 

[1] Department of Chemical Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Colaba, Mumbai 400005, India

关键词: Cytochrome P450cam;    Substrate binding;    Substrate access channel;    Steady-state fluorescence;    Picosecond time-resolved fluorescence;    Conformational change;   

DOI  :  10.1016/S0014-5793(00)01745-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The binding of camphor to cytochrome P450_cam has been investigated by steady-state and time-resolved tryptophan fluorescence spectroscopy to obtain information on the substrate access channel. The fluorescence quenching experiments show that some of the tryptophan residues undergo changes in their local environment on camphor binding. The time-resolved fluorescence decay profile gives four lifetime components in the range from 99 ps to 4.5 ns. The shortest lifetime component assigned to W42 lies close to the proposed camphor access channel. The results show that the fluorescence of W42 is greatly affected on binding of camphor, and supports dynamic fluctuations involved in the passage of camphor through the access channel as proposed earlier on the basis of crystallographic, molecular dynamics simulation and site-directed mutagenesis studies.

【 授权许可】

Unknown