title: Ribose 5-phosphate glycation reduces cytochrome c respiratory activity and membrane affinity.
abstract: Spontaneous glycation of bovine heart cytochrome c (cyt c) by the sugar ribose 5-phosphate (R5P) weakens the ability of the heme protein to transfer electrons in the respiratory pathway and to bind to membranes. Trypsin fragmentation studies suggest the preferential sites of glycation include Lys72 and Lys87/88 of a cationic patch involved in the association of the protein with its respiratory chain partners and with cardiolipin-containing membranes. Reaction of bovine cyt c with R5P (50 mM) for 8 h modified the protein in a manner that weakened its ability to transfer electrons to cytochrome oxidase by 60%. An 18 h treatment with R5P decreased bovine cyt c's binding affinity with cardiolipin-containing liposomes by an estimated 8-fold. A similar weaker binding of glycated cyt c was observed with mitoplasts. The reversal of the effects of R5P on membrane binding by ATP further supports an A-site modification. A significant decrease in the rate of spin state change for ferro-cyt c, thought to be due to cardiolipin insertion disrupting the coordination of Met to heme, was found for the R5P-treated cyt c. This change occurred to a greater extent than what can be explained by the permanent attachment of the protein to the liposome. Turbidity changes resulting from the multilamellar liposome fusion that is readily promoted by cyt c binding were not seen for the R5P-glycated cyt c samples. Collectively, these results demonstrate the negative impact that R5P glycation can have on critical electron transfer and membrane association functions of cyt c.
authors.affiliation: Department of Chemistry and Biochemistry, Middlebury College, Middlebury, Vermont 05753, United States.
authors.forename: Gordon J
authors.initials: GJ
authors.lastname: Hildick-Smith
authors.forename: Michael C
authors.initials: MC
authors.lastname: Downey
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Gretebeck
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Gersten
authors.forename: Roger K
authors.initials: RK
authors.lastname: Sandwick
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002308:Cardiolipins / Q000378:metabolism; D002417:Cattle; D045304:Cytochromes c / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000378:metabolism; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D008081:Liposomes; D008239:Lysine / Q000737:chemistry / Q000378:metabolism; D008561:Membrane Fusion; D051336:Mitochondrial Membranes / Q000201:enzymology / Q000378:metabolism*; D009391:Nephelometry and Turbidimetry; D015388:Organelles / Q000201:enzymology / Q000378:metabolism; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D012267:Ribosemonophosphates / Q000737:chemistry / Q000378:metabolism*; D012995:Solubility
chemical_list: D002308:Cardiolipins; D005656:Fungal Proteins; D008081:Liposomes; D010446:Peptide Fragments; D012267:Ribosemonophosphates; C031626:ribose-5-phosphate; D000255:Adenosine Triphosphate; D045304:Cytochromes c; D003576:Electron Transport Complex IV; D008239:Lysine
references.citation: Protein Sci. 1993 Jun;2(6):966-76
references.pmid: 8391357
references.citation: Ann N Y Acad Sci. 2005 Jun;1043:85-96
references.pmid: 16037226
references.citation: Biochem J. 1969 Oct;115(1):65-75
references.pmid: 5388143
references.citation: J Biol Chem. 1992 Nov 5;267(31):22243-8
references.pmid: 1331048
references.citation: Carbohydr Res. 2007 Dec 10;342(17):2575-92
references.pmid: 17850774
references.citation: Biochim Biophys Acta. 1959 Sep;35:130-40
references.pmid: 14421038
references.citation: Biochemistry. 1995 Feb 28;34(8):2686-93
references.pmid: 7873551
references.citation: J Biol Chem. 1994 Jan 21;269(3):1770-4
references.pmid: 8294426
references.citation: J Biol Chem. 1978 Jan 10;253(1):149-59
references.pmid: 201616
references.citation: J Biol Chem. 1996 Aug 2;271(31):18379-86
references.pmid: 8702481
references.citation: J Biol Chem. 2000 May 26;275(21):16127-33
references.pmid: 10821864
references.citation: J Am Chem Soc. 2007 Jan 24;129(3):504-5
references.pmid: 17227009
references.citation: J Biol Chem. 2005 Oct 14;280(41):34709-17
references.pmid: 16012169
references.citation: J Biol Chem. 1961 Aug;236:2342-6
references.pmid: 13756114
references.citation: Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):2800-5
references.pmid: 11880631
references.citation: J Biol Chem. 1980 May 25;255(10):4732-9
references.pmid: 6246081
references.citation: Carbohydr Res. 2010 Nov 22;345(17):2499-506
references.pmid: 20933223
references.citation: J Biol Chem. 2010 Mar 26;285(13):9317-9321
references.pmid: 20139083
references.citation: Biochem J. 1991 Nov 1;279 ( Pt 3):781-6
references.pmid: 1659388
references.citation: Proc Natl Acad Sci U S A. 1981 Mar;78(3):1456-60
references.pmid: 6262802
references.citation: J Biol Chem. 1972 Oct 25;247(20):6375-83
references.pmid: 4342598
references.citation: Biophys Chem. 2003 Sep;105(2-3):743-55
references.pmid: 14499930
references.citation: Free Radic Biol Med. 2009 Jun 1;46(11):1439-53
references.pmid: 19285551
references.citation: Biochem J. 1988 Jun 1;252(2):349-55
references.pmid: 2843168
references.citation: J Biol Inorg Chem. 2010 Jun;15(5):689-700
references.pmid: 20238133
references.citation: J Biol Chem. 1995 Feb 17;270(7):3197-202
references.pmid: 7852404
references.citation: Nat Cell Biol. 2008 Dec;10(12):1477-83
references.pmid: 19029908
references.citation: Arch Biochem Biophys. 1991 Jul;288(1):112-7
references.pmid: 1654817
references.citation: Mol Cell Biochem. 2010 Aug;341(1-2):149-57
references.pmid: 20352475
references.citation: Biochem J. 2007 Oct 15;407(2):179-87
references.pmid: 17614790
references.citation: J Biol Chem. 2002 Mar 15;277(11):8822-6
references.pmid: 11781329
references.citation: J Biol Chem. 1973 Jan 25;248(2):528-33
references.pmid: 4346335
references.citation: Mol Cells. 2006 Oct 31;22(2):220-7
references.pmid: 17085975
references.citation: Biochem Cell Biol. 2000;78(6):675-81
references.pmid: 11206578
references.citation: Biochemistry. 2006 Apr 18;45(15):4998-5009
references.pmid: 16605268
references.citation: J Biol Chem. 1978 Sep 10;253(17):6045-53
references.pmid: 210168
references.citation: Biochem Biophys Res Commun. 1997 Sep 8;238(1):140-2
references.pmid: 9299467
references.citation: Biochemistry. 2008 Jul 1;47(26):6928-35
references.pmid: 18540683