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Crystallographic Analyses of Site-Directed Mutants of the Photosynthetic Reaction Center from Rhodobacter sphaeroides

Chirino, Arthur J. and Lous, Erik J. and Huber, Martina and Allen, James P. and Schenck, Craig C. and Paddock, Mark L. and Feher, George and Rees, Douglas C. (1994) Crystallographic Analyses of Site-Directed Mutants of the Photosynthetic Reaction Center from Rhodobacter sphaeroides. Biochemistry, 33 (15). pp. 4584-4593. ISSN 0006-2960. https://resolver.caltech.edu/CaltechAUTHORS:20150205-141324993

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Abstract

Seven site-directed mutants of the bacterial photosynthetic reaction center (RC) from the 2.4.1 and WS 231 wild-type strains of Rhodobacter sphaeroides have been crystallized and their X-ray diffraction analyzed to resolutions between 3.0 and 4.0 Å. The mutations can be divided into four distinct categories: (1) mutations altering cofactor composition that affect electron transfer and quantum yield, His M202 → Leu (M202HL), His L173 → Leu (L173HL), and Leu M214 → His (M214LH); (2) a mutation in the proposed pathway of electron transfer altering electron-transfer kinetics, Tyr M210 → Phe (M210YF); (3) a mutation around the non-heme iron resulting in an iron-less reaction center, His M219 → Cys (M219HC); and (4) mutations around the secondary electron acceptor, a ubiquinone, affecting proton transfer and quinone turnover, Glu L212 → Gin (L212EQ) and Asp L213 → Asn (L213DN). Residues L173 and M202 are within bonding distance of the respective magnesiums of the two bacteriochlorophylls of the BChl special pair, while M214 is close to the bacteriopheophytin on the active A branch of the RC. The L173HL and M202HL crystal structures show that the respective bacteriochlorophylls are replaced with bacteriopheophytins (i.e., loss of magnesium) without significant structural perturbations to the surrounding main-chain or side-chain atoms. In the M214LH mutant, the bacteriopheophytin has been replaced by a bacteriochlorophyll, and the side chain of His M214 is within ligand distance of the magnesium. The M210YF, L212EQ, and L213DN mutants show no significant tertiary structure changes near the mutation sites. The M219HC diffraction data indicate that the overall tertiary structure of the reaction center is maintained in the absence of the non-heme iron.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/bi00181a020 DOIArticle
http://pubs.acs.org/doi/abs/10.1021/bi00181a020PublisherArticle
ORCID:
AuthorORCID
Rees, Douglas C.0000-0003-4073-1185
Contact Email Address:phoebe@caltech.edu
Additional Information:© 1994 American Chemical Society. Received September 21, 1993; Revised Manuscript Received February 7, 1994. This work was supported by the National Institutes of Health (GM45162 to D.C.R., GM41300 to J.P.A., GM13191 to G.F., GM41637 to M.L.P., and R.C.D.A. GM00536 and GM38214/48254 to C.C.S.). E.J.L. was supported by the European Molecular Biology Organisation and M.H. by a Forschungsstipendium of the Deutsche Forschungsgemeinschaft. Use of the X-ray data collection facilities at UCLA and the Cray YMP at the National Science Foundation-supported San Diego Supercomputer Center is gratefully acknowledged.
Funders:
Funding AgencyGrant Number
NIHGM45162
NIHGM41300
NIHGM13191
NIHGM41637
NIHGM00536
NIHGM38214/48254
European Molecular Biology Organization (EMBO)UNSPECIFIED
Deutsche Forschungsgemeinschaft (DFG)UNSPECIFIED
Issue or Number:15
Record Number:CaltechAUTHORS:20150205-141324993
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150205-141324993
Official Citation:Crystallographic Analyses of Site-Directed Mutants of the Photosynthetic Reaction Center from Rhodobacter sphaeroides Arthur J. Chirino, Erik J. Lous, Martina Huber, James P. Allen, Craig C. Schenck, Mark L. Paddock, George Feher, and Douglas C. Rees Biochemistry 1994 33 (15), 4584-4593
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:54447
Collection:CaltechAUTHORS
Deposited By: SWORD User
Deposited On:08 Feb 2015 02:55
Last Modified:03 Oct 2019 07:57

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