Purification and molecular characterization of a secretory transglutarninase from coagulating gland of the rat

Seitz, Jürgen; Keppler, Claudia; Hüntemann, S; Rausch, Ulrich; Aumüller, Gerhard

doi:10.1016/0167-4838(91)99002-a

Cited by 34 publications

(20 citation statements)

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“…The high mannose-to-galactose ratio, unusual for complex type glycans, supports the lectin assay data indicating the occurrence of high mannose structures. Moreover, since lectin binding assays ruled out the occurrence of O-linked structures, the presence of N-acetylgalactosamine together with that of myo-inositol and the high content of mannose residues suggests the existence of a glycosylphosphatidylinositol anchor in rat CGS TGase, as previously hypothesized (15).…”

Section: Post-translational Modificationsmentioning

confidence: 69%

“…TGase isolated from the secretion of rat coagulating gland has been reported as a single highly glycosylated polypeptide chain (M r ϭ 65,000) with a lipid anchor retained during the enzyme apocrine secretion process (15,28,29). The only data reported so far on rat CGS TGase amino acid sequence derive from homology studies between different molecular forms of TGase and the cDNA-derived primary structure of the major androgen-dependent secretory protein of rat coagulating gland (DP1) (11).…”

Section: Discussionmentioning

confidence: 99%

“…The basic biochemical characteristics of the enzyme and the formation of glutamine-lysine intermolecular cross-links of the vesicular secretory proteins were described first by Lorand and WilliamsAshman (14). Recently, rat prostate TGase was reported to be a highly glycosylated polypeptide chain (M r ϭ 65,000) and to possess a lipid anchor that is retained during secretion (15).…”

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confidence: 99%

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Transglutaminase from Rat Coagulating Gland Secretion

Esposito

Pucci

Amoresano³

et al. 1996

Journal of Biological Chemistry

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Structural and biochemical characteristics of transglutaminase purified by a rapid chromatographic procedure from the rat coagulating gland (anterior prostate) secretion are reported. Fast atom bombardment mapping and automated Edman degradation experiments allowed us to verify that at least 85% of the entire transglutaminase amino acid sequence is identical to that derived from the cDNA of the major androgen-dependent rat prostate protein called DP1. The enzyme was found NH 2 terminally blocked and largely posttranslationally modified, since the presence of N-linked oligosaccharides, as well as of complex lipidic structures, was observed. Mass spectral analysis showed that Asn-408 and -488 are the glycosylated sites, the N-linked structures identified belonging to both high-mannose and complex type glycans. The presence of myo-inositol, of glycerol bound fatty acids, and the high content of mannose residues, are in agreement with previous observations suggesting that a lipid anchor is bound to coagulating gland secretion transglutaminase. Furthermore, two tightly bound calcium ions per molecule of enzyme were detected. Finally, a strong stimulation of the enzyme activity in vitro by both SDS and a variety of phosphatidic acids was observed. The reported structural and functional peculiarities should definitively lead to consider the prostate enzyme as a new member (type IV) of the transglutaminase family.

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Section: Post-translational Modificationsmentioning

confidence: 69%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Transglutaminase from Rat Coagulating Gland Secretion

Esposito

Pucci

Amoresano³

et al. 1996

Journal of Biological Chemistry

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“…Unlike a secretory transglutaminase from rat liver [36], no change in the PI value of pTGase was noticed as the degree of purification increased. The 1 % (by vol.)…”

Section: Characterization Of Ptgasementioning

confidence: 97%

Purification and characterization of a novel transglutaminase from filarial nematode Brugia malayi

Singh

Mehta

1994

European Journal of Biochemistry

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A transglutaminase (pTGase) was purified from filarial nematode, Brugia malayi. The steps used for purification were thermoprecipitation, ammonium sulfate precipitation, gel filtration on Superose 12 HR 10/30, ion-exchange chromatography on a Mono-Q column and further gel filtration on Superose 12 HR 10/30. The last step yielded an electrophoretically homogenous enzyme protein with 2200-fold purification and a reproducible yield of approximately 20%. The purified enzyme had a molecular mass of 56 kDa, specific activity of 2.2.5 U/mg protein and an isoelectric point of 7.2. The enzyme was active in the basic pH range with an optimum activity at pH 8.5. The pTGase activity was Ca2+-dependent and was inhibited by ammonia, primary amines, EDTA, and -SH group blocking reagents. The enzyme activity was also inhibited by high salt (NaC1 and KC1) concentrations, detergents, metal ions, and organic solvents. Ampholine (pH 6-8) at 1 % (by vol.) caused about 20% inhibition of pTGase activity but at 3% (by vol.) the inhibition increased up to 80%. Similarly, the micromolar concentrations of GTP inhibited the enzyme activity only moderately but at millimolar concentration a significant inhibition was observed. The stability of the pTGase was not affected by 0.1% SDS or other physical parameters such as freezing and thawing. Further, the pTGase was found to be highly thermostable (stable at 60°C for several hours) with optimum activity observed at 55 "C. The distinct substrate specificity, unique N-terminal sequence along with the other physico-chemical properties studied, suggested that pTGase is a novel member of transglutaminase family.Transglutaminases (TGase, EC 2.3.2.13) are a family of Ca2+-dependent enzymes that catalyze the formation of isopeptide bonds between protein-bound glutamine residues and primary amine groups (such as polyamines), resulting in proteidprotein cross-links or amine/protein conjugates [l, 21. Recently, a transglutaminase from a bacterial source was isolated that, unlike most known transglutaminases, did not require Ca2+ for its catalytic activity [3].Transglutaminases are widely distributed enzymes among mammalian tissues and organs [2, 41 and

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“…Original magnification 200. tissues have been isolated and characterized on their genetic and biochemical properties, such as TGase 1 (keratinocyte TGase, particulate TGase, TGase K), TGase 2 (tissue TGase, liver TGase, cytosolic TGase, TGase C), TGase 3 (epidermal TGase, TGase E), TGase 4 (prostate TGase) and Factor XIII. Each enzyme has been found to be expressed organ-specifically and associated with a variety of biological processes (Folk and Cole, 1966;Takagi and Doolittle, 1974;Thatcher, 1989;Kim et al, 1990;Seitz et al, 1991).…”

Section: Five Different Types Of Tgases From the Mammalian 181mentioning

confidence: 99%

Isolation and characterization of brain-specific transglutaminases from rat

Kwak

Kim²,

Kim³

et al. 1998

Exp Mol Med

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The relevance of transglutaminases with neural function and several disorders has been emphasized recently. Especially, many polypeptides associated with neurodegenerative diseases are suggested to be putative transglutaminase substrates such as amyloid protein of Altzheimer's disease, microtubule-associated proteins and neurofilaments, etc. In addition, the CAG repeated gene products with probable polyglutamine tract, putative transglutami-nase substrates, were identified in several neuro-degenerative disorders. However, the identity of the brain transglutaminase has not been confirmed, because of enzymic stability and low activity. In the present experiment, we have isolated brain-specific transglutaminases, designated as TGase NI and TGase NII, which are different from other types of transglutaminases in respects of molecular weights (mw. 45 kDa, 29 kDa respectively), substrate affinity, elution profile on ion-exchange chromatography, sensitivity to proteases and ethanol, and immuno-logical properties. The enzymes were localized specifically in the brain tissues but not in the liver tissue. And neural cells such as pheochromocytoma cell, glioma cell, primary neuronal and glial cells were shown to be enriched with TGase NI and TGase NII. The possible biological roles of the enzymes were discussed not only on the aspect of crosslinking activity but also of signal transducing capacity of the enzyme in the brain. K e y w o r d s : TGase NI, TGaase NII, brain, rat, localization

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Purification and molecular characterization of a secretory transglutarninase from coagulating gland of the rat

Cited by 34 publications

References 0 publications

Transglutaminase from Rat Coagulating Gland Secretion

Transglutaminase from Rat Coagulating Gland Secretion

Purification and characterization of a novel transglutaminase from filarial nematode Brugia malayi

Isolation and characterization of brain-specific transglutaminases from rat

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