Amino acid sequence of guinea pig liver transglutaminase from its cDNA sequence

Ikura, Koji; Nasu, Takaaki; Yokota, Hiroyuki; Tsuchiya, Yoichi; Sasaki, Ryuzo; Chiba, Hideo

doi:10.1021/bi00408a035

Cited by 185 publications

(116 citation statements)

References 49 publications

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“…We revealed that processing of TGase 1 is very specific, but by unknown protease(s) giving 20 times higher activity than intact form (Kim et al, 1995;. TGase 2 is an ubiquitous enzyme expressed in every cells and tissues (Ikura et al, 1988;Gentile et al, 1991). In the human epidermis, TGase 2 is expressed mostly in the supra basal layer and dermal layer which suggests it might be related with cell proliferation and extracellular matrix formation (Aeschilimann and Paulsson, 1991).…”

Section: Introductionmentioning

confidence: 83%

Calpain inhibitors reduce the cornified cell envelope formation by inhibiting proteolytic processing of transglutaminase 1

Kim

Bae²

1998

Exp Mol Med

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Calpain I ( -calpain) and II (m-calpain) are well known calcium-activated neutral cysteine proteases. Many reports have shown that activation of calpain is related to cataract formation, neuronal degeneration, blood clotting, ischemic injuries, muscular dystrophy and cornified cell envelope (CE) formation. Here, we report that insoluble CE formation was reduced after treatment with calpain I inhibitor (N-acetyl-leucylleucyl-norleucinal) on normal human epidermal keratinocytes (NHEK), whereas serine and thiol protease inhibitors had no effect on the reduction of CE. When NHEK cells were confluent, keratinocytes were treated with various concentrations (0.5 M -0.5 mM) of calpain I inhibitor or serine and thiol protease inhibitors under calcium induced differentiation. Insoluble CE formation was reduced about 90 % in the 50 M calpain inhibitor I treated group by day 9 of culture, whereas insoluble CE was reduced only 10 % in the same condition. Interestingly TGase activity was blocked by 90 % in the 0.5 mM calpain inhibitor treated group within 72 h, whereas TGase activity was retained by 80 % in the 0.5 mM serine protease inhibitor treated group at 7 day treatment. Therefore it can be suggested that cysteine protease calpains might be responsible for the activation of the TGase 1 enzyme to complete insoluble CE formation during epidermal differentiation.

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Section: Introductionmentioning

confidence: 83%

Calpain inhibitors reduce the cornified cell envelope formation by inhibiting proteolytic processing of transglutaminase 1

Kim

Bae²

1998

Exp Mol Med

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“…In the region marked c in Fig. 4 2), which contains the transglutaminase active site in XIIIa (24) and LTG (25). The catalytic cysteine of the active sites of the transglutaminases and the corresponding alanine in P4.2 (see arrow in large box in Fig.…”

Section: Methodsmentioning

confidence: 99%

“…4, b) (24,26,27) and LTG (25), especially around their active sites. XIIIa, the final component in the coagulation pathway, plays an important role in the stabilization of fibrin clots by covalently crosslinking fibrin monomers through y-glutamyl-E-lysine bridges and by preventing proteolysis (35).…”

Section: Methodsmentioning

confidence: 99%

Molecular cloning of human protein 4.2: a major component of the erythrocyte membrane.

Sung¹,

Chien²,

Chang³

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

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Protein 4.2 (P4.2) comprises -5% of the protein mass of human erythrocyte (RBC) membranes. Anemia occurs in patients with RBCs deficient in P4.2, suggesting a role for this protein in maintaining RBC stability and integrity. We now report the molecular cloning and characterization of human RBC P4.2 cDNAs. By immunoscreening a human reticulocyte cDNA library and by using the polymerase chain reaction, two cDNA sequences of 2.4 and 2.5 kilobases (kb) were obtained. These cDNAs differ only by a 90-base-pair insert in the longer isoform located three codons downstream from the putative initiation site. The 2.4-and 2.5-kb cDNAs predict proteins of -77 and =80 kDa, respectively, and the authenticity was confimed by sequence identity with 46 amino acids of three cyanogen bromide-cleaved peptides of P4. Subcloning and Sequence Analysis. cDNA inserts from positive phage clones were subcloned into pBS(+) plasmids (Stratagene). Unidirectional deletion clones were generated by using BAL-31 exonuclease (13), and cDNA fragments were sequenced with T3 and T7 primers by the dideoxynucleotide chain-termination method (14). Sequence analysis and GenBank data base searches were performed by IBI Pustell sequence analysis software (International Biotechnologies). 5'-End Extension of cDNA. Three oligonucleotides were prepared in a technique based on the polymerase chain reaction (PCR) to synthesize the missing 5' sequence of the partial cDNA clone: pl was composed of nucleotides (nt) 7-23 of clone 7 (c.7); p2 was complementary to nt 36-52 of c.7 plus an EcoRI restriction site at its 5' end; p3 was composed of the EcoRI polylinker and the poly(dC) originally used for the first-strand cDNA synthesis when the library was constructed. The sequences of pl, p2, and p3 were, respectively, 5'-dTGAGGATGCTGTGTTCC-3', 5'-dTCGAAlJCGTACTC-CATGCGCTGAG-3', and 5'-dGCGGLAAlTCCCCCCCCCC-CCCC-3', with the EcoRI sites underlined. p2 and p3 were used as PCR primers, and the reticulocyte cDNA library (5 ,u with 106 phages per pl) was used as a template. The reaction product was electrophoresed and stained with ethidium bromide. The major band was excised and subcloned into pGEM 3zf plasmids (Promega). From >500 transformants, 24 colonies were randomly chosen to make minipreparations of plasmid DNA, of which 80%o were positive when hybridized with 32P-labeled pl. tTo whom reprint requests should be addressed. IThe sequences reported in this paper have been deposited in the GenBank data base (accession nos. M30646 and M30647). 955The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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“…These enzymes catalyze protein crosslinking reactions via the formation of e-(c-glutamyl)lysine bonds between the c-carboxyl group of a Gln residue in one polypeptide chain and the e-amino group of a Lys residue in a second polypeptide chain. Well-documented examples of TG are plasma factor XIIIa [2], keratinocyte TG [3], epidermal TG [4], tissue TG [5], and prostatic TG [6]. Recent findings have shown that, apart from their protein modifying capabilities, tissue TG is also able to function as a component of the signal-transducing G protein complex [7].…”

mentioning

confidence: 99%

Molecular characterization of a novel nuclear transglutaminase that is expressed during starfish embryogenesis

Sugino¹,

Terakawa²,

Yamasaki³

et al. 2002

Eur J Biochem

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We report the constitution and molecular characterization of a novel transglutaminase (EC 2.3.2.13) that starts to accumulate specifically in the nucleus in the starfish (Asterina pectinifera) embryo after progression through the early blastula stage. The cDNA for the nuclear transglutaminase was cloned and the cDNA-deduced sequence defines a single open reading frame encoding a protein with 737 amino acids and a predicted molecular mass of 83 kDa. A comparison of this transglutaminase with other members of the gene family revealed an overall sequence identity of 33-41%. A special sequence feature of this transglutaminase, which is not found in other transglutaminases, is the presence of nuclear localization signal-like sequences in the N-terminal region.Microinjection of hybrid constructs that encode the N-terminal segment fused to reporter proteins into the germinal vesicle of an oocyte produced chimeric proteins by transcription-coupled translation. It was found that the N-terminal segment alone was sufficient to effect nuclear accumulation of an otherwise cytoplasmic protein. These results suggest that the nuclear accumulation of the transglutaminase may play an important role in nuclear remodeling during early starfish embryogenesis.Keywords: transglutaminase; nucleus; starfish; embryo; cloning.The class of enzymes that are commonly referred to as transglutaminases (TG) (EC 2.3. Recent findings have shown that, apart from their protein modifying capabilities, tissue TG is also able to function as a component of the signal-transducing G protein complex [7]. The cDNA of G ha , involved in the transmission of adrenergic stimuli, is identical to that of tissue TG of human endothelial cells [7]. Tissue TG is localized mainly in the cytosol, but detectable tissue TG expression has been reported in the nucleus [8][9][10]. However, TG activity in the nucleus and the mechanisms of its translocation is not well understood, and nucleus-specific TG has not been reported.It is accepted that many proteins are able to cross nuclear membranes and accumulate against gradients to concentrate in the nucleus [11,12]. The nuclear translocation of proteins via the nuclear pore complex is dependent on a nuclear localization signal in the protein, which is rich in basic amino acids and may be bipartite [13][14][15]. The functional assays of such nuclear localization signals are usually based on the ability of a signal to confer nuclear localization to an otherwise non-nuclear protein.The present paper describes the occurrence of a novel TG that is localized exclusively in the nucleus of starfish (Asterina pectinifera) embryonic cells and is designated nuclear TG (nTG). The amino-acid sequence derived from the cDNA sequence contains putative nuclear localization signals [15] in the N-terminal region. We demonstrate here that the N-terminal region promotes the nuclear accumulation of an otherwise cytoplasmic protein, namely pyruvate kinase (PK), in the A. pectinifera oocyte system. This finding suggests that nuclear localization sig...

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Amino acid sequence of guinea pig liver transglutaminase from its cDNA sequence

Cited by 185 publications

References 49 publications

Calpain inhibitors reduce the cornified cell envelope formation by inhibiting proteolytic processing of transglutaminase 1

Calpain inhibitors reduce the cornified cell envelope formation by inhibiting proteolytic processing of transglutaminase 1

Molecular cloning of human protein 4.2: a major component of the erythrocyte membrane.

Molecular characterization of a novel nuclear transglutaminase that is expressed during starfish embryogenesis

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