Cloning, heterologous expression, and characterization of recombinant class II cytochromes c from Rhodopseudomonas palustris
Abstract
The cytochrome (cyt) c′, cyt c_(556), and cyt c_2 genes from Rhodopseudomonas palustris have been cloned; recombinant cyt c′ and cyt c_(556) have been expressed, purified, and characterized. Unlike mitochondrial cyt c, these two proteins are structurally similar to cyt b_(562), in which the heme is embedded in a four-helix bundle. The hemes in both recombinant proteins form covalent thioether links to two Cys residues. UV/vis spectra of the Fe^(II) and Fe^(III) states of the recombinant cyts are identical with those of the corresponding native proteins. Equilibrium unfolding measurements in guanidine hydrochloride solutions confirm that native Fe^(II)-cyt c_(556) is more stable than the corresponding state of Fe^(III)-cyt c_(556) (ΔΔG_f°=22 kJ/mol).
Additional Information
© 2002 Elsevier. Received 16 July 2002; received in revised form 9 September 2002; accepted 19 September 2002. The authors thank Linda Thöny-Meyer for the gift of plasmid pEC86, and A. Grant Mauk for the gift of plasmid pBPCYC1. This work was supported by NSF (MCB 9974477), NIH (GM16424, J.H.R.; GM020316-03, M.A.M.; GM07616, J.G.L), the Ralph M. Parsons Foundation (J.C.L.), and the Arnold and Mabel Beckman Foundation.Additional details
- Eprint ID
- 56981
- DOI
- 10.1016/S0304-4165(02)00437-3
- Resolver ID
- CaltechAUTHORS:20150424-155358251
- MCB-9974477
- NSF
- GM16424
- NIH
- GM020316-03
- NIH
- GM07616
- NIH
- Ralph M. Parsons Foundation
- Arnold and Mabel Beckman Foundation
- Created
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2015-04-24Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field