Comparison of Substrate Specificities of Transglutaminases Using Synthetic Peptides as Acyl donors

Ohtsuka, Tomoko; Ota, Masafumi; Nio, Noriki; Motoki, Masao

doi:10.1271/bbb.64.2608

Cited by 87 publications

(55 citation statements)

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“…2 ) are concentrated in the front vestibule of the active site cleft. Interestingly, an experiment concerning the substrate specificity for the acyl donor of MTG demonstrated that synthetic peptides containing amino acid residues other than Gly and positively charged residues at the N-terminal side of Gln are good substrates of MTG, whereas MTG drastically loses its catalytic efficiency with the peptides containing residues other than Gly at the C-terminal side of Gln (45). These results suggest that the N-terminal side of Gln within the acyl donor binds to the front vestibule of the active site cleft of MTG and that MTG requires an acyl donor with large conformational flexibility and small side chains on the C-terminal side of Gln, to avoid steric hindrance with the enzyme itself.…”

Section: Resultsmentioning

confidence: 99%

Crystal Structure of Microbial Transglutaminase fromStreptoverticillium mobaraense

Kashiwagi

Yokoyama

Ishikawa

et al. 2002

Journal of Biological Chemistry

230

224

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The crystal structure of a microbial transglutaminase from Streptoverticillium mobaraense has been determined at 2.4 Å resolution. The protein folds into a platelike shape, and has one deep cleft at the edge of the molecule. Its overall structure is completely different from that of the factor XIII-like transglutaminase, which possesses a cysteine protease-like catalytic triad. superimpose well on the catalytic triad "Cys-HisAsp" of the factor XIII-like transglutaminase, in this order. The secondary structure frameworks around these residues are also similar to each other. These results imply that both transglutaminases are related by convergent evolution; however, the microbial transglutaminase has developed a novel catalytic mechanism specialized for the cross-linking reaction. The structure accounts well for the catalytic mechanism, in which Asp 255 is considered to be enzymatically essential, as well as for the causes of the higher reaction rate, the broader substrate specificity, and the lower deamidation activity of this enzyme.Transglutaminase (TGase 1 ; protein-glutamine ␥-glutamyltransferase, EC 2.3.2.13) catalyzes an acyl transfer reaction in which the ␥-carboxyamide groups of peptide-bound glutamine residues act as the acyl donors. The most common acyl acceptors of TGase are the ⑀-amino groups of lysine residues within peptides or the primary amino groups of some naturally occurring polyamines (1, 2). When lysine residues in proteins serve as acyl acceptors, intermolecular or intramolecular ⑀-(␥-glutamyl)lysine bonds are formed, resulting in the polymerization of proteins.TGases are widely distributed in various organisms, including vertebrates (3-7), invertebrates (8, 9), mollusks (10), plants (11), and microorganisms (12). Among these TGases, the human blood coagulation factor XIII has been most characterized (13)(14)(15)(16)(17)(18). By catalyzing the cross-linking between fibrin molecules, factor XIII forms fibrin clots for hemostasis and heals a wound. The crystal structure of human factor XIII has been determined, revealing that it consists of four domains with a cysteine protease-like active site (19 -22). Many TGases are homologous to human factor XIII and share the common feature of Ca 2ϩ -dependent catalytic activity (3-8). A tissue-type TGase from red sea bream liver (fish-derived TGase (FTG)) is an example of such factor XIII-like TGases and shows 33% sequence homology to human factor XIII (7). The crystal structure of FTG has also been determined (23). The overall and active site structures of FTG are essentially similar to those of human factor XIII.A microbial TGase (MTG) has been isolated from the culture medium of Streptoverticillium sp. S-8112 (24), which has been identified as a variant of Sv. mobaraense. This enzyme is the first TGase obtained from a nonmammalian source. Thus far, few TGases have been identified from microorganisms, particularly from Streptoverticillium species (25). Although the physiological role of MTG is still unknown, this protein is secreted from the cytoplas...

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Section: Resultsmentioning

confidence: 99%

Crystal Structure of Microbial Transglutaminase fromStreptoverticillium mobaraense

Kashiwagi

Yokoyama

Ishikawa

et al. 2002

Journal of Biological Chemistry

230

224

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“…The biochemical functions of microbial TGase (MTGase) from Streptomyces mobaraense (BCRC 12165, ATCC 29032; Streptoverticillium mobaraense, Streptomyces ladakanum, and Streptoverticillium ladakanum) have been studied extensively (5)(6)(7). This Ca 2ϩ -independent enzyme is secreted as a zymogen with an additional prosequence that consists of 45 amino acid residues (DNGAGEETKSYAETYRLTADDVANI-NALNESAPAASSAGPSFRAP) at the N terminus.…”

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confidence: 99%

Crystal Structure and Inhibition Studies of Transglutaminase from Streptomyces mobaraense

Yang¹,

Chang

Wang

et al. 2011

Journal of Biological Chemistry

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The crystal structure of the microbial transglutaminase (MTGase) zymogen from Streptomyces mobaraense has been determined at 1.9-Å resolution using the molecular replacement method based on the crystal structure of the mature MTGase. The overall structure of this zymogen is similar to that of the mature form, consisting of a single disk-like domain with a deep active cleft at the edge of the molecule.

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“…In designing new lipid substrates for MTG, an MTG-reactive Gln-containing peptidyl substrate was conjugated with fatty acids. Previous studies of peptide screening [11] and docking simulations [12] have indicated that N-terminal hydrophobic residues from the reactive Gln residue was critical for substrate recognition by MTG, and we found that MTG recognizes a seven amino acid sequence (GGGSLLQG) at the C terminus of recombinant proteins. [13] With this in mind, we synthesized lipid-modified peptides (C14-1, C16-1, C18-1, Scheme 1 a) in which the N terminus of the MTG-reactive Gln donor peptide was attached to diverse long-chain fatty acids (myristic acid, palmitic acid and stearic acid) using solid-phase peptide synthesis.…”

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confidence: 57%

Protein Lipidation Catalyzed by Microbial Transglutaminase

Abe

Goto

Kamiya

2011

Chemistry A European J

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Fattening up: Artificially constructed lipid–protein conjugates can be used to control protein activity and localization at either biological or artificial membrane interfaces. Here, we report a facile technique for the lipid modification of proteins catalyzed by microbial transglutaminase (MTG). MTG‐mediated protein lipidation proceeded efficiently without perturbing protein functionality (see figure).

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Comparison of Substrate Specificities of Transglutaminases Using Synthetic Peptides as Acyl donors

Cited by 87 publications

References 1 publication

Crystal Structure of Microbial Transglutaminase fromStreptoverticillium mobaraense

Crystal Structure of Microbial Transglutaminase fromStreptoverticillium mobaraense

Crystal Structure and Inhibition Studies of Transglutaminase from Streptomyces mobaraense

Protein Lipidation Catalyzed by Microbial Transglutaminase

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