High-resolution mass spectrometry analysis of protein oxidations and resultant loss of function.
MS, with or without pre-analysis peptide fractionation, can be used to decipher the residues on proteins where oxidative modifications caused by peroxynitrite, singlet oxygen or electrophilic lipids have occurred. Peroxynitrite nitrates tyrosine and tryptophan residues on the surface of actin. Singlet oxygen, formed by the interaction of UVA light with tryptophan, can oxidize neighbouring cysteine, histidine, methionine, tyrosine and tryptophan residues. Dose-response inactivation by 4HNE (4-hydroxynonenal) of hBAT (human bile acid CoA:amino acid N-acyltransferase) and CKBB (cytosolic brain isoform of creatine kinase) is associated with site-specific modifications. FT-ICR (Fourier-transform ion cyclotron resonance)-MS using nanoLC (nano-liquid chromatography)-ESI (electrospray ionization)-MS or direct-infusion ESI-MS with gas-phase fractionation identified 14 4HNE adducts on hBAT and 17 on CKBB respectively. At 4HNE concentrations in the physiological range, one member of the catalytic triad of hBAT (His362) was modified; for CKBB, although all four residues in the active site that were modifiable by 4HNE were ultimately modified, only one, Cys283, occurred at physiological concentrations of 4HNE. These results suggest that future in vivo studies should carefully assess the critical sites that are modified rather than using antibodies that do not distinguish between different modified sites.
Barnes
Stephen
S
Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, USA. sbarnes@uab.edu
Shonsey
Erin M
EM
Eliuk
Shannon M
SM
Stella
David
D
Barrett
Kerri
K
Srivastava
Om P
OP
Kim
Helen
H
Renfrow
Matthew B
MB
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D052276:Creatine Kinase, BB Form / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D017550:Spectroscopy, Fourier Transform Infrared; D038203:alpha-Crystallin B Chain / Q000235:genetics / Q000378:metabolism
D000447:Aldehydes; C059215:CRYAB protein, human; D015853:Cysteine Proteinase Inhibitors; D038203:alpha-Crystallin B Chain; D000217:Acyltransferases; C026953:bile acid-CoA amino acid N-acyltransferase; D052276:Creatine Kinase, BB Form; C027576:4-hydroxy-2-nonenal
J Am Soc Mass Spectrom. 2004 Aug;15(8):1136-47
15276160
Nat Rev Drug Discov. 2007 Aug;6(8):662-80
17667957
Nat Rev Mol Cell Biol. 2007 Sep;8(9):722-8
17700625
Biochemistry. 2005 Sep 13;44(36):12245-52
16142923
J Biol Chem. 2002 Dec 6;277(49):47270-5
12239217
Anal Chem. 1988 Oct 15;60(20):2299-301
3239801
Proteomics. 2001 Jan;1(1):108-17
11680890
J Mol Med (Berl). 2002 Aug;80(8):463-74
12185447
Chem Res Toxicol. 2003 Jul;16(7):901-11
12870893
Glycoconj J. 2006 Jul;23(5-6):291-302
16897173
Free Radic Biol Med. 2003 Dec 1;35(11):1417-30
14642389
Mol Vis. 2008 Feb 01;14:234-48
18334940
Biochemistry. 2003 Aug 26;42(33):9906-14
12924939
PLoS One. 2008 Jan 30;3(1):e1529
18231610
Methods Enzymol. 1994;233:229-40
8015460
J Biol Chem. 1997 Sep 5;272(36):22472-80
9278398
Eur J Biochem. 2004 Jul;271(13):2841-52
15206949
Science. 1989 Oct 6;246(4926):64-71
2675315
J Mol Biol. 2008 Feb 29;376(4):997-1007
18190928
J Photochem Photobiol B. 2001 Oct;63(1-3):114-25
11684458
Free Radic Biol Med. 2008 May 1;44(9):1700-11
18242196
Nat Methods. 2005 Jan;2(1):17-25
15789031
Free Radic Res. 1996 Aug;25(2):149-59
8885333
Free Radic Biol Med. 2000 May 1;28(9):1370-8
10924856
J Lipid Res. 2008 Feb;49(2):282-94
17965457
Chem Res Toxicol. 1996 Mar;9(2):390-6
8839040
J Biol Chem. 2003 May 30;278(22):19587-90
12679331
Pediatr Res. 2000 Apr;47(4 Pt 1):492-4
10759156
Biochimie. 2005 Aug;87(8):673-85
15927344
Methods Enzymol. 2005;405:139-71
16413314
Nature. 1985 Jan 24-30;313(6000):277-84
2578615
Cell Mol Life Sci. 2007 Nov;64(22):2863-86
17717629
Methods Enzymol. 2005;405:116-38
16413313
Methods Mol Biol. 2007;371:349-91
17634592
Chem Res Toxicol. 2007 Sep;20(9):1260-8
17696488
Biochim Biophys Acta. 2005 Jan 17;1703(2):93-109
15680218
Trends Genet. 2006 Oct;22(10):545-54
16908088
Mutat Res. 2007 May 1;618(1-2):102-15
17374386
Free Radic Biol Med. 2004 Sep 1;37(5):671-81
15288124
Annu Rev Biophys Biomol Struct. 2005;34:267-94
15869391
J Biol Chem. 2003 Mar 7;278(10):8380-4
12475975
J Biol Chem. 1992 Jul 5;267(19):13425-33
1320020
J Nutr. 2003 Nov;133(11 Suppl 1):3773S-3777S
14608113
Annu Rev Biochem. 2008;77:755-76
18173371