The Enzymatic Deamidation of Proteins

Mycek, Mary J.; Waelsch, Heinrich

doi:10.1016/s0021-9258(18)64499-0

Cited by 71 publications

(17 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…That shifted ratio suggested either an increased direct hydrolytic deamidation pathway that led to the formation of α-Glu product or a decreased concentration of γ-Glu product due to the existence of intermediates in the transamidation of these modified Gln sites (Figure b). Enzymatic deamidation of Gln residues is followed by transamidation when the deamidation reaction is mediated by TGs and lysine residues are available. ,, This TG-mediated reaction results in the formation of an intra- or intermolecular ε-(γ-glutamyl)lysine covalent bond. , …”

Section: Resultsmentioning

confidence: 99%

“…Enzymatic deamidation of Gln residues is followed by transamidation when the deamidation reaction is mediated by TGs and lysine residues are available. 11,30,31 This TG-mediated reaction results in the formation of an intra-or intermolecular ε-(γ-glutamyl)lysine covalent bond. 8,13 To identify the presence of intramolecular cross-linked transamidation in brain proteins, we searched for trypticdigested peptides that met the following three criteria: 11,12 (1) display a mass loss of 17.03 Da due to the loss of ammonia as a result of the formation of the intramolecular ε-(γ-glutamyl)lysine bond, (2) contain a miscleaved Lys that is not digested by trypsin due to its participation in the ε-(γ-glutamyl)lysine bond, and (3) contain a Gln located at the vicinity of the miscleaved Lys residue that participates in the formation of the ε-(γ-glutamyl)lysine bond.…”

Section: Analytical Chemistrymentioning

confidence: 99%

See 1 more Smart Citation

Characterization of Glutamine Deamidation by Long-Length Electrostatic Repulsion-Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry (LERLIC-MS/MS) in Shotgun Proteomics

et al. 2016

View full text Add to dashboard Cite

Deamidation of glutamine (Gln) residues is a spontaneous or enzymatic process with significant implications in aging and human pathology. Although some methods are available to identify the γ/α-glutamyl products of deamidation, none of these methods allows the characterization of this post-translational modification (PTM) from complex biological samples by shotgun proteomics. Here we present LERLIC-MS/MS, a chromatographic strategy that uses a long (50 cm) anion-exchange capillary column operating in the electrostatic repulsion-hydrophilic interaction mode (ERLIC) and coupled directly to tandem mass spectrometry (MS/MS) for proteome analysis in a single injection. Profiling of soluble extracts of brain tissues by LERLIC-MS/MS distinguished for the first time γ/α-glutamyl isomers of deamidation, encountering a 1.7 γ/α-glutamyl ratio for most Gln deamidation products. A detailed analysis of any deviation from that observed ratio allowed the identification of transglutaminase-mediated γ-glutamyl isomers as intermediate products of transamidation. Furthermore, LERLIC-MS/MS was able to simultaneously separate Gln and asparagine (Asn) deamidation products even for those peptides showing multiple deamidated proteoforms. The characterization of Asn deamidated residues by LERLIC-MS/MS also uncovered novel PIMT (protein L-isoaspartyl methyltransferase) substrate proteins in human brain tissues that deviated from the expected 3:1 isoAsp/Asp ratio. Taken together, our results demonstrate that LERLIC-MS/MS can be used to perform an in-depth study of protein deamidation on a global proteome scale. This new strategy should help to elucidate the biological implications of deamidation in aging and disease conditions.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Analytical Chemistrymentioning

confidence: 99%

Characterization of Glutamine Deamidation by Long-Length Electrostatic Repulsion-Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry (LERLIC-MS/MS) in Shotgun Proteomics

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Transglutaminase was prepared according to the procedure of Clarke et al (1959) from the soluble fraction of guinea pig liver homogenate. This enzyme preparation showed a specific activity of 250 units/mg of protein when assayed by means of the N-Z Amine substrate of Mycek and Waelsch (1960).…”

Section: Methodsmentioning

confidence: 99%

Peptidoglutaminase. Enzymes for selective deamidation of γ-amide of peptide-bound glutamine

Kikuchi¹,

Hayashida²,

Nakano³

et al. 1971

Biochemistry

View full text Add to dashboard Cite

“…Functional output of proteins is primarily regulated by a diverse array of posttranslational modifications, such as phosphorylation, ubiquitination, sumoylation, ISGylation, acetylation, methylation, and deamidation (1)(2)(3)(4). Although reported more than half a century ago (5), protein deamidation has been largely regarded as a side effect of protein "aging" or functional decay (6). Thus, our understanding of protein deamidation is rudimentary at best.…”

Section: Introductionmentioning

confidence: 99%

Antiviral activity of a purine synthesis enzyme reveals a key role of deamidation in regulating protein nuclear import

Zhao

et al. 2019

Sci. Adv.

View full text Add to dashboard Cite

Protein nuclear translocation is highly regulated and crucial for diverse biological processes. However, our understanding concerning protein nuclear import is incomplete. Here we report that a cellular purine synthesis enzyme inhibits protein nuclear import via deamidation. Employing human Kaposi’s sarcoma-associated herpesvirus (KSHV) to probe the role of protein deamidation, we identified a purine synthesis enzyme, phosphoribosylformylglycinamidine synthetase (PFAS) that inhibits KSHV transcriptional activation. PFAS deamidates the replication transactivator (RTA), a transcription factor crucial for KSHV lytic replication. Mechanistically, deamidation of two asparagines flanking a positively charged nuclear localization signal impaired the binding of RTA to an importin β subunit, thus diminishing RTA nuclear localization and transcriptional activation. Finally, RTA proteins of all gamma herpesviruses appear to be regulated by PFAS-mediated deamidation. These findings uncover an unexpected function of a metabolic enzyme in restricting viral replication and a key role of deamidation in regulating protein nuclear import.

show abstract

The Enzymatic Deamidation of Proteins

Cited by 71 publications

References 10 publications

Characterization of Glutamine Deamidation by Long-Length Electrostatic Repulsion-Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry (LERLIC-MS/MS) in Shotgun Proteomics

Characterization of Glutamine Deamidation by Long-Length Electrostatic Repulsion-Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry (LERLIC-MS/MS) in Shotgun Proteomics

Peptidoglutaminase. Enzymes for selective deamidation of γ-amide of peptide-bound glutamine

Antiviral activity of a purine synthesis enzyme reveals a key role of deamidation in regulating protein nuclear import

Contact Info

Product

Resources

About