Characterization of the Metal-Binding Site of Bovine Growth Hormone through Site-Specific Metal-Catalyzed Oxidation and High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Hovorka, Susan W.; Williams, Todd D.; Schöneich, Christian

doi:10.1006/abio.2001.5447

Cited by 34 publications

(10 citation statements)

References 19 publications

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“…Protein oxidation and fragmentation by MCO takes place in close proximity to the metal binding site as a consequence of extreme reactivity and the short life of the hydroxyl radical generated. Thus, MCO has been utilized to identify the metal binding sites of a number of proteins (32–35). …”

Section: Introductionmentioning

confidence: 99%

Distinct oxidative cleavage and modification of bovine [Cu– Zn]-SOD by an ascorbic acid/Cu(II) system: Identification of novel copper binding site on SOD molecule

Uehara

Luo

Aryal

et al. 2016

Free Radical Biology and Medicine

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We investigated the combined effect of ascorbate and copper [Asc/Cu(II)] on the integrity of bovine [Cu-Zn]-superoxide dismutase (bSOD1) as a model system to study the metal catalyzed oxidation (MCO) and fragmentation of proteins. We found Asc/Cu(II) mediates specific cleavage of bSOD1 and generates 12.5 and 3.2 kDa fragments in addition to oxidation/carbonylation of the protein. The effect of other tested transition metals, a metal chelator, and hydrogen peroxide on the cleavage and oxidation indicated that binding of copper to a previously unknown site on SOD1 is responsible for the Asc/Cu(II) specific cleavage and oxidation. We utilized tandem mass spectrometry to identify the specific cleavage sites of Asc/Cu(II)-treated bSOD1. Analyses of tryptic- and AspN-peptides have demonstrated the cleavage to occur at Gly31 with peptide bond breakage with Thr30 and Ser32 through diamide and α-amidation pathways, respectively. The three-dimensional structure of bSOD1 reveals the imidazole ring of His19 localized within 5 Angstrom from the α-carbon of Gly31 providing a structural basis that copper ion, most likely coordinated by His19, catalyzes the specific cleavage reaction.

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Section: Introductionmentioning

confidence: 99%

Distinct oxidative cleavage and modification of bovine [Cu– Zn]-SOD by an ascorbic acid/Cu(II) system: Identification of novel copper binding site on SOD molecule

Uehara

Luo

Aryal

et al. 2016

Free Radical Biology and Medicine

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“…These methods use radicals (e.g. • OH) to modify solvent‐exposed4–8 or metal‐bound amino acids 9–20. After oxidation, modifications to amino acid side chains are usually identified by digesting the protein of interest with a proteolytic enzyme and subjecting the modified peptides to tandem MS (MS/MS) analyses.…”

Section: Introductionmentioning

confidence: 99%

Correct identification of oxidized histidine residues using electron‐transfer dissociation

2009

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Oxidative modification to the side chain of histidine can noticeably change the collision-induced dissociation (CID) pathways of peptides containing this oxidized residue. In cases where an oxidized peptide consists of two or more isomers differing only in the site of modification, oxidation to histidine usually causes the other oxidized sites to be mis-assigned in CID spectra. These spectral mis-assignments can sometimes be avoided by using multiple stages of MS/MS (MS n ) or via specially-optimized liquid chromatographic separation conditions. In this manuscript, we demonstrate that these mis-assignments can be more readily and easily avoided by using electrontransfer dissociation (ETD) to dissociate the oxidized peptides. Furthermore, we find that the relative insensitivity of ETD to side chain chemistry allows the extent of oxidative modification to be determined readily for peptide isomers having more than one site of oxidation. The current results along with previous studies of oxidized peptides suggest that ETD is probably a better technique than CID for obtaining correct sequence and modification information for oxidized peptides.

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“…These methods use radicals (e.g. • OH) to modify solvent-exposed [1][2][3][4][5][6][7][8][9][10] or metal-bound amino acids [11][12][13][14][15][16][17][18]. Tandem MS (MS/MS), typically in conjunction with proteolytic digestion, is then used to identify oxidatively modified residues, and information about protein structure is then derived.…”

Section: Introductionmentioning

confidence: 99%

The effect of histidine oxidation on the dissociation patterns of peptide ions

Bridgewater

Rapole

et al. 2007

J. Am. Soc. Mass Spectrom.

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Oxidative modifications to amino acid side chains can change the dissociation pathways of peptide ions, but these variations are most commonly observed when cysteine and methionine residues are oxidized. In this work we describe the very noticeable effect that oxidation of histidine residues can have on the dissociation patterns of peptide ions containing this residue. A common product ion spectral feature of doubly-charged tryptic peptides is enhanced cleavage at the C-terminal side of histidine residues. This preferential cleavage arises due to the unique acid/base character of the imidazole side chain that initiates cleavage of a proximal peptide bond for ions in which the number of protons does not exceed the number of basic residues. We demonstrate here that this enhanced cleavage is eliminated when histidine is oxidized to 2-oxo-histidine because the proton affinity and nucleophilicity of the imidazole side chain are lowered. Furthermore, we find that oxidation of histidine to 2-oxo-histidine can cause the mis-assignment of oxidized residues when more than one oxidized isomer is simultaneously subjected to tandem mass spectrometry (MS/MS). These spectral misinterpretations can be avoided usually by using multiple stages of MS/MS (MS n ) or via speciallyoptimized liquid chromatographic separation conditions. When these approaches are not accessible or do not work, N-terminal derivatization with sulfobenzoic acid avoids the problem of mistakenly assigning oxidized residues.

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Characterization of the Metal-Binding Site of Bovine Growth Hormone through Site-Specific Metal-Catalyzed Oxidation and High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Cited by 34 publications

References 19 publications

Distinct oxidative cleavage and modification of bovine [Cu– Zn]-SOD by an ascorbic acid/Cu(II) system: Identification of novel copper binding site on SOD molecule

Distinct oxidative cleavage and modification of bovine [Cu– Zn]-SOD by an ascorbic acid/Cu(II) system: Identification of novel copper binding site on SOD molecule

Correct identification of oxidized histidine residues using electron‐transfer dissociation

The effect of histidine oxidation on the dissociation patterns of peptide ions

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