Amino‐acid‐sequence Determination and Biological Activity of Cytin, a Naturally Occurring Specific Chymotrypsin Inhibitor from the Leech <i>Theromyzon tessulatum</i>

Chopin, Vincent; Bilfinger, Thomas V.; Stefano, George B.; Matias, Isabel

doi:10.1111/j.1432-1033.1997.t01-1-00733.x

Cited by 16 publications

(18 citation statements)

References 24 publications

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“…By contrast, sequence comparisons were carried out for theromin with the four different serine protease inhibitors isolated from the leech T. tessulatum: cytin, therin, therostasin, and tessulin (5,29,(41)(42)(43). This revealed that two of the four (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…The proportion of activated cells was determined as described elsewhere (40). Activated cells not only change their conformation in response to a pharmacological stimulus, but also become mobile and capable of phagocytosis and secrete various signaling molecules (40,41).…”

Section: Methodsmentioning

confidence: 99%

“…Immunocyte Assay-Cells were separated by a standard FicollHypaque method as described elsewhere in detail (40,41). They were washed three times in RPMI medium (RPMI, 25 mM Hepes, Grand Island Biological Co., New York, NY).…”

Section: Methodsmentioning

confidence: 99%

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Theromin, a Novel Leech Thrombin Inhibitor

Salzet¹,

Chopin²,

Baert³

et al. 2000

Journal of Biological Chemistry

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We purified the most potent thrombin inhibitor described to date from the rhynchobdellid leech Theromyzon tessulatum. Designated theromin, it was purified to apparent homogeneity by gel permeation and anion exchange chromatography followed by two reverse-phase steps of high performance liquid chromatography. The primary sequence of theromin (a homodimer of 67 amino acid residues including 16 cysteine residues) was determined by a combination of reduction and s-␤-pyridylethylation, Edman degradation, trypsin enzymatic digestion, and matrix-assisted laser desorption mass spectrometry measurement. Theromin exhibits no sequence homology with any other thrombin inhibitors. Furthermore, theromin significantly diminishes, in a dose-dependent manner, the level of human granulocyte and monocyte activation induced by lipopolysaccharides. In summary, this potent thrombin inhibitor promises to have high biomedical significance.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Theromin, a Novel Leech Thrombin Inhibitor

Salzet¹,

Chopin²,

Baert³

et al. 2000

Journal of Biological Chemistry

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confidence: 57%

“…We now report on yet another inhibitor, designated tessulin. We also demonstrate that this molecule modulates [in conjunction with other T. tessulatum serine-protease inhibitors, such as therin [22], cytin [23] and thero-min (thrombin inhibitor isolated from T. tessulatum ; Matias, I., Malecha, J. and Salzet, M., unpublished results)] the activity of human immunocytes, in a higher manner to aprotinin, a serineprotease inhibitor used to diminish the diffuse inflammatory response associated with major surgery [24,25]. Thus, leeches represent a reservoir of serine-protease inhibitors which are so essential for a parasitic life style involving fluid acquisition when clotting processes are present in the host.…”

mentioning

confidence: 59%

Amino‐acid‐sequence determination and biological activity of tessulin, a naturally occurring trypsin‐chymotrypsin inhibitor isolated from the leech Theromyzon tessulatum

Chopin¹,

Stefano

1998

European Journal of Biochemistry

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We purified a new trypsinϪchymotrypsin inhibitor, designated tessulin, from the rhynchobdellid leech Theromyzon tessulatum. This 9-kDa peptide was purified to apparent homogeneity by gel-permeation and anion-exchange chromatographies followed by reverse-phase HPLC. The structure of tessulin was determined by reduction, S-β-pyridylethylation, trypsin digestion, automated Edman degradation and matrix-assisted laser desorption mass spectrometry (m/z 8985 Da). The 81-amino-acid peptide possesses 16 cysteines and exhibits a 16% sequence similarity with antistasin-type inhibitors. Tessulin inhibits trypsin (K i 1 pM) and chymotrypsin (K i 150 pM) and exhibits no activity with thrombin, factor Xa, cathepsin G and elastase. This is the first trypsinϪchymotrypsin inhibitor isolated from leeches that does not inhibit elastase or cathepsin G, except for cytin and therin. Furthermore, tessulin, in conjunction with other serine-protease inhibitors isolated from Theromyzon (therin, theromin), significantly diminishes the level of human granulocyte and monocyte activation induced by lipopolysaccharides (10 µg). The combined level of inhibition is higher than that of aprotinin, another serine-protease inhibitor used biomedically. Thus, tessulin may be clinically significant in reducing inflammatory events.Keywords : protease inhibitors; trypsin-chymotrypsin inhibitor.In numerous pathological processes, the crucial role played by proteases is becoming more apparent. This critical role creates a need for the development of non-toxic protease inhibitors for in vivo clinical applications. Because of the specific recognition by proteases of defined amino acid sequences, it is possible to inhibit these enzymes when they are involved in pathological processes [1]. The involvement of proteolytic enzymes in a number of degenerative diseases, e.g., pancreatitis [zymogenic forms of proteolytic enzymes (trypsin 1 and 2, chymotrypsin A [2, 3] and elastase [4])], is becoming more evident as is the need for curtailing this activity in uncalled-for situations [5Ϫ7]. In this regard, aprotinin, a 6.5-kDa trypsin and kallikrein inhibitor isolated from bovine organs [8] has been used for more than three decades as an intensive care drug for acute pancreatitis [9], demonstrating the significance of this class of enzyme.In hematophageous leeches, animals that must escape hostimmune surveillance, studies on serine-protease inhibitors have yielded significant findings relating to the clinical importance of these enzyme inhibitors. Of two groups of serine-protease [19]. However, some of these inhibitors exhibit different amino acid sequences and inhibitory activities; their three-dimensional structures share the same structural motif with the leech antihemostatic protein (L.A.P.) [20]. Interestingly, their mechanisms of action and epitopes important for binding to their respective targets are distinct as well [20], suggesting that through evolution different strategies have emerged to deal with a similar problem.Among the second group of serine-protea...

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Evidence for an Annelid Neuroendocrine System

Vieau

Breton

2004

Cell Signalling in Prokaryotes and Lower Metazoa

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Hormonal processes along with enzymatic processing similar to that found in vertebrates occur in annelids. Amino acid sequence determination of annelids precursor gene products reveals the presence of the respective peptides that exhibit high sequence identity to their mammalian counterparts. Furthermore, these neuropeptides exert similar physiological function in annelids than the ones found in vertebrates. In this respect, the high conservation in course of evolution of these molecules families reflects their importance. Nevertheless, some specific neuropeptides to annelids or invertebrates have also been in these animals.

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Amino‐acid‐sequence Determination and Biological Activity of Cytin, a Naturally Occurring Specific Chymotrypsin Inhibitor from the Leech Theromyzon tessulatum

Cited by 16 publications

References 24 publications

Theromin, a Novel Leech Thrombin Inhibitor

Theromin, a Novel Leech Thrombin Inhibitor

Amino‐acid‐sequence determination and biological activity of tessulin, a naturally occurring trypsin‐chymotrypsin inhibitor isolated from the leech Theromyzon tessulatum

Evidence for an Annelid Neuroendocrine System

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