Identification of glycinin in vivo as a polyamine-conjugated protein via a γ-glutamyl linkage

Kang, Hyeog; Lee, Seung Gwan; Cho, Young Dong

doi:10.1042/bj3320467

Cited by 8 publications

(2 citation statements)

References 41 publications

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“…As a consequence, formation of a protein polymer (de novo direct polymerization) [ 6 ], stabilization of non-covalent assemblies by introducing covalent bonds between adjacent subunits—known as enzymatic “spot welding”—or conformational restriction of a folded polypeptide takes place [ 1 , 7 , 8 , 9 ]. Apart from their cross-linking activity, this class of enzymes has been reported to carry out other enzymatic activities including protein phosphorylation [ 10 ], glutamine deamidation [ 11 ], amine incorporation [ 12 ] and esterification [ 13 ]. Abnormal TG cross-linking activity is implicated in different diseases, including fibrosis [ 14 , 15 , 16 ], pancreatic cancer [ 17 , 18 ] and celiac disease [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Mass Spectrometric Identification of a Novel Factor XIIIa Cross-Linking Site in Fibrinogen

Semkova

Hsuan

2021

Proteomes

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Transglutaminases are a class of enzymes that catalyze the formation of a protein:protein cross-link between a lysine and a glutamine residue. These cross-links play important roles in diverse biological processes. Analysis of cross-linking sites in target proteins is required to elucidate their molecular action on target protein function and the molecular specificity of different transglutaminase isozymes. Mass-spectrometry using settings designed for linear peptide analysis and software designed for the analysis of disulfide bridges and chemical cross-links have previously been employed to identify transglutaminase cross-linking sites in proteins. As no control peptide with which to assess and improve the mass spectrometric analysis of TG cross-linked proteins was available, we developed a method for the enzymatic synthesis of a well-defined transglutaminase cross-linked peptide pair that mimics a predicted tryptic digestion product of collagen I. We then used this model peptide to determine optimal score thresholds for correct peptide identification from y- and b-ion series of fragments produced by collision-induced dissociation. We employed these settings in an analysis of fibrinogen cross-linked by the transglutaminase Factor XIIIa. This approach resulted in identification of a novel cross-linked peptide in the gamma subunit. We discuss the difference in behavior of ions derived from different cross-linked peptide sequences and the consequent demand for a more tailored mass spectrometry approach for cross-linked peptide identification compared to that routinely used for linear peptide analysis.

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Section: Introductionmentioning

confidence: 99%

Mass Spectrometric Identification of a Novel Factor XIIIa Cross-Linking Site in Fibrinogen

Semkova

Hsuan

2021

Proteomes

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“…The presence of the AtPng1p mRNA, even though low expressed, in all tissues, growth stages, and light conditions suggests that AtPng1p gene product could be a constitutive enzyme. Lilley et al (1998) as well as Falcone et al (1993) and Kang et al (1998) reported the occurrence of TGase activities in several organs of the same plant.…”

Section: Recombinant Atpng1p Protein Conjugates Spermine To Glutamyl mentioning

confidence: 99%

AtPng1p. The First Plant Transglutaminase

Mea

Caparrós-Ruiz²,

Claparols³

et al. 2004

Plant Physiology

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Studies have revealed in plant chloroplasts, mitochondria, cell walls, and cytoplasm the existence of transglutaminase (TGase) activities, similar to those known in animals and prokaryotes having mainly structural roles, but no protein has been associated to this type of activity in plants. A recent computational analysis has shown in Arabidopsis the presence of a gene, AtPng1p, which encodes a putative N-glycanase. AtPng1p contains the Cys-His-Asp triad present in the TGase catalytic domain. AtPng1p is a single gene expressed ubiquitously in the plant but at low levels in all light-assayed conditions. The recombinant AtPng1p protein could be immuno-detected using animal TGase antibodies. Furthermore, western-blot analysis using antibodies raised against the recombinant AtPng1p protein have lead to its detection in microsomal fraction. The purified protein links polyamines-spermine (Spm) . spermidine (Spd) . putrescine (Put)-and biotin-cadaverine to dimethylcasein in a calcium-dependent manner. Analyses of the g-glutamyl-derivatives revealed that the formation of covalent linkages between proteins and polyamines occurs via the transamidation of g-glutamyl residues of the substrate, confirming that the AtPng1p gene product acts as a TGase. The Ca 21-and GTP-dependent cross-linking activity of the AtPng1p protein can be visualized by the polymerization of bovine serum albumine, obtained, like the commercial TGase, at basic pH and in the presence of dithiotreitol. To our knowledge, this is the first reported plant protein, characterized at molecular level, showing TGase activity, as all its parameters analyzed so far agree with those typically exhibited by the animal TGases.Transglutaminases (TGases; E.C. 2.3.2.13) catalyze protein cross-linking by the interaction of an acyl acceptor glutamyl residue and an amine donor, a lysyl residue, of the same or another protein, or the formation of a link between a protein and a free primary amine, like a polyamine (PA). The terminal aminogroups of PAs conjugate one or two glutamyl residues giving rise either to mono-(g-glutamyl)-polyamines (mono-PAs) or bis-(g-glutamyl)-polyamines (bis-PAs; Folk, 1980). The posttranslational cross-linking of proteins is one of the essential physiological processes involved in the stabilization of tissue and cellular matrices. High molecular protein networks can be generated by TGase catalysis when cross-links are formed between Gln and either Lys residues or PAs (bis-PA derivatives) thus forming bridges between different proteins. At present, both in eukaryotes and prokaryotes several TGases have been described exhibiting a large number of functions (Aeschlimann et al., 1998;Griffin et al., 2002;Lorand and Graham, 2003). Eight distinct TGase isoenzymes have been identified in different animal tissues, and some of these have been purified and characterized at molecular level (Aeschlimann et al., 1998;Griffin et al., 2002;Lorand and Graham, 2003).TGases are classified in three structural families: (1) the papain-like TGases, (2) the protein di...

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