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Ruffling of Metalloporphyrins Bound to IsdG And IsdI, Two Heme Degrading Enzymes in Staphylococcus Aureus

Journal Article · · J. Biol. Chem. 283:30957,2008
OSTI ID:953086
IsdG and IsdI are paralogous proteins that are intracellular components of a complex heme uptake system in Staphylococcus aureus. IsdG and IsdI were shown previously to reductively degrade hemin. Crystal structures of the apoproteins show that these proteins belong to a newly identified heme degradation family distinct from canonical eukaryotic and prokaryotic heme oxygenases. Here we report the crystal structures of an inactive N7A variant of IsdG in complex with Fe{sup 3+}-protoporphyrin IX (IsdG-hemin) and of IsdI in complex with cobalt protoporphyrin IX (IsdI-CoPPIX) to 1.8 {angstrom} or better resolution. These structures show that the metalloporphyrins are buried into similar deep clefts such that the propionic acids form salt bridges to two Arg residues. His{sup 77} (IsdG) or His{sup 76} (IsdI), a critical residue required for activity, is coordinated to the Fe{sup 3+} or Co{sup 3+} atoms, respectively. The bound porphyrin rings form extensive steric interactions in the binding cleft such that the rings are highly distorted from the plane. This distortion is best described as ruffled and places the {beta}- and {delta}-meso carbons proximal to the distal oxygen-binding site. In the IsdG-hemin structure, Fe{sup 3+} is pentacoordinate, and the distal side is occluded by the side chain of Ile{sup 55}. However, in the structure of IsdI-CoPPIX, the distal side of the CoPPIX accommodates a chloride ion in a cavity formed through a conformational change in Ile{sup 55}. The chloride ion participates in a hydrogen bond to the side chain amide of Asn{sup 6}. Together the structures suggest a reaction mechanism in which a reactive peroxide intermediate proceeds with nucleophilic oxidation at the {beta}- or {delta}-meso carbon of the hemin.
Research Organization:
Stanford Linear Accelerator Center (SLAC)
Sponsoring Organization:
USDOE
DOE Contract Number:
AC02-76SF00515
OSTI ID:
953086
Report Number(s):
SLAC-REPRINT-2009-198
Journal Information:
J. Biol. Chem. 283:30957,2008, Journal Name: J. Biol. Chem. 283:30957,2008 Journal Issue: 45 Vol. 283; ISSN JBCHA3; ISSN 0021-9258
Country of Publication:
United States
Language:
English

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