Document Detail

Plastocyanin-cytochrome f interactions: the influence of hydrophobic patch mutations studied by NMR spectroscopy.
MedLine Citation:
PMID:  12501198     Owner:  NLM     Status:  MEDLINE    
Transient complex formation between plastocyanin from Prochlorothrix hollandica and cytochrome f from Phormidium laminosum was investigated using nuclear magnetic resonance (NMR) spectroscopy. Binding curves derived from NMR titrations at 10 mM ionic strength reveal a 1:1 stoichiometry and a binding constant of 6 (+/-2) x 10(3) M(-1) for complex formation, 1 order of magnitude larger than that for the physiological plastocyanin-cytochrome f complex from Ph. laminosum. Chemical-shift perturbation mapping indicates that the hydrophobic patch of plastocyanin is involved in the complex interface. When the unusual hydrophobic patch residues of P. hollandica plastocyanin were reverted to the conserved residues found in most other plastocyanins (Y12G/P14L), the binding constant for the interaction with cytochrome f was unaffected. However, the chemical shift perturbation map was considerably different, and the size of the average perturbation decreased by 40%. The complexes of both the wild-type and double mutant plastocyanin with cytochrome f were sensitive to ionic strength, contrary to the physiological complex. The possible implications of these findings for the mechanism of transient complex formation are discussed.
Peter B Crowley; Nadejda Vintonenko; George S Bullerjahn; Marcellus Ubbink
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Biochemistry     Volume:  41     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  2002 Dec 
Date Detail:
Created Date:  2002-12-26     Completed Date:  2003-02-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  15698-705     Citation Subset:  IM    
Gorlaeus Laboratories, Leiden Institute of Chemistry, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
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MeSH Terms
Binding Sites / genetics
Conserved Sequence / genetics
Cyanobacteria / enzymology,  genetics
Cytochromes / chemistry*,  genetics
Cytochromes f
Glycine / genetics
Histidine / chemistry
Leucine / genetics
Macromolecular Substances
Mutagenesis, Site-Directed*
Nuclear Magnetic Resonance, Biomolecular* / methods
Plastocyanin / chemistry*,  genetics
Prochlorothrix / chemistry,  genetics
Proline / genetics
Tyrosine / genetics
Reg. No./Substance:
0/Cytochromes; 0/Macromolecular Substances; 0/Protons; 147-85-3/Proline; 55520-40-6/Tyrosine; 56-40-6/Glycine; 61-90-5/Leucine; 71-00-1/Histidine; 9014-09-9/Plastocyanin; 9035-46-5/Cytochromes f

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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