Interactions of Streptomyces Serine-Protease Inhibitors with Streptomyces griseus Metalloendopeptidase II

Kajiwara, Kenta; Fujita, Akira; Tsuyuki, H.; Kumazaki, Takashi; Ishii, Shin‐ichi

doi:10.1093/oxfordjournals.jbchem.a123584

Cited by 16 publications

(14 citation statements)

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“…The removal of both amino acids resulted in full depletion of inhibitory activity. We assume that SSI and other SSI-like metalloprotease inhibitors such as SgiA use, alike SSTI, this inhibition site, despite the fact that the exchange of amino acids within the subtilisin binding loop may alter the dissociation constant by one order of magnitude [5]. Our conclusion was supported by former studies with SSI-like Streptomyces cinnamoneus subsp.…”

Section: Discussionsupporting

confidence: 81%

“…Moreover, SSI proteins strongly affect activity of the Streptomyces griseus metalloprotease II (SGMPII, SgmA, griselysin, http://www.uniprot.org/uniprot/B1W035/http://www.uniprot.org/uniprot/Q83WH1), which was thought to compete with subtilisin BPN’ for the same inhibition site . However, substitution or deletion of SSI‐Met104 and adjacent amino acids could not decisively break the inhibition of SGMPII, and dissociation constants were at most altered by one order of magnitude . In contrast, the Streptomyces caespitosus neutral protease inhibitor (ScNPI, http://www.uniprot.org/uniprot/Q9FDS0) for the metalloprotease ScNP (snapalysin, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/24/77.html, http://www.uniprot.org/uniprot/P56406), originally obtained from the undefined Streptomyces isolate I‐355, was completely inactivated by the exchange of ScNPI‐Tyr61 for alanine that is located two residues downstream from the N‐terminal cysteine and opposite to the subtilisin‐affine methionine (ScNPI‐Met99) .…”

Section: Introductionmentioning

confidence: 99%

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The N‐terminal peptide of the transglutaminase‐activating metalloprotease inhibitor from Streptomyces mobaraensis accommodates both inhibition and glutamine cross‐linking sites

et al. 2019

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Streptomyces mobaraensis is a key player for the industrial production of the protein cross‐linking enzyme microbial transglutaminase (MTG). Extra‐cellular activation of MTG by the transglutaminase‐activating metalloprotease (TAMP) is regulated by the TAMP inhibitory protein SSTI that belongs to the large Streptomyces subtilisin inhibitor (SSI) family. Despite decades of SSI research, the binding site for metalloproteases such as TAMP remained elusive in most of the SSI proteins. Moreover, SSTI is a MTG substrate, and the preferred glutamine residues for SSTI cross‐linking are not determined. To address both issues, that is, determination of the TAMP and the MTG glutamine binding sites, SSTI was modified by distinct point mutations as well as elongation or truncation of the N‐terminal peptide by six and three residues respectively. Structural integrity of the mutants was verified by the determination of protein melting points and supported by unimpaired subtilisin inhibitory activity. While exchange of single amino acids could not disrupt decisively the SSTI TAMP interaction, the N‐terminally shortened variants clearly indicated the highly conserved Leu40‐Tyr41 as binding motif for TAMP. Moreover, enzymatic biotinylation revealed that an adjacent glutamine pair, upstream from Leu40‐Tyr41 in the SSTI precursor protein, is the preferred binding site of MTG. This extension peptide disturbs the interaction with TAMP. The structure of SSTI was furthermore determined by X‐ray crystallography. While no structural data could be obtained for the N‐terminal peptide due to flexibility, the core structure starting from Tyr41 could be determined and analysed, which superposes well with SSI‐family proteins. Enzymes Chymotrypsin, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/21/1.html; griselysin (SGMPII, SgmA), EC3.4.24.27; snapalysin (ScNP), http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/24/77.html; streptogrisin‐A (SGPA), http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/21/80.html; streptogrisin‐B (SGPB), http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/21/81.html; subtilisin BPN’, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/21/62.html; transglutaminase, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC2/3/2/13.html; transglutaminase‐activating metalloprotease (TAMP), EC3.4.‐.‐; tri‐/tetrapeptidyl aminopeptidase, EC3.4.11.‐; trypsin, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/21/4.html. Databases The atomic coordinates and structure factors (PDB 6I0I) have been deposited in the Protein Data Bank (http://www.rcsb.org).

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Section: Discussionsupporting

confidence: 81%

Section: Introductionmentioning

confidence: 99%

The N‐terminal peptide of the transglutaminase‐activating metalloprotease inhibitor from Streptomyces mobaraensis accommodates both inhibition and glutamine cross‐linking sites

et al. 2019

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“…Nevertheless, it is interesting that a molecule structurally related to the inhibitors of one class of proteases (i.e., serine proteases) regulates a member of another class of proteases (i.e., MMPs). This is not, however, unprecedented: several serine protease inhibitors, including ␣-macroglobulins (34), the Streptomyces subtilisin family of inhibitors (35), and the amyloid precursor protein (36), have been reported to inhibit metalloproteinases. Further studies on the interaction between RECK and MMP-9 may yield new insights into how such cross-class proteaseprotease inhibitor interactions occur.…”

Section: Discussionmentioning

confidence: 99%

Regulation of matrix metalloproteinase-9 and inhibition of tumor invasion by the membrane-anchored glycoprotein RECK

Takahashi

Sheng

Horan

et al. 1998

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

437

544

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A human fibroblast cDNA expression library was screened for cDNA clones giving rise to f lat colonies when transfected into v-Ki-ras-transformed NIH 3T3 cells. One such gene, RECK, encodes a membrane-anchored glycoprotein of about 110 kDa with multiple epidermal growth factor-like repeats and serine-protease inhibitor-like domains. While RECK mRNA is expressed in various human tissues and untransformed cells, it is undetectable in tumor-derived cell lines and oncogenically transformed cells. Restored expression of RECK in malignant cells resulted in suppression of invasive activity with concomitant decrease in the secretion of matrix metalloproteinase-9 (MMP-9), a key enzyme involved in tumor invasion and metastasis. Moreover, purified RECK protein was found to bind to, and inhibit the proteolytic activity of, MMP-9. Thus, RECK may link oncogenic signals to tumor invasion and metastasis.Mutations of ras protooncogenes are found in a large variety of human tumors (1). It has been well established that Ras proteins are essential components in various intracellular signaling pathways involved in regulating gene expression and several other aspects of cellular behavior (2). Therefore, it is now important to find targets for these signals relevant to the expression of the malignant phenotype to understand the mechanism of cell transformation and to develop means to cure or prevent cancers.To this end, we have been isolating and characterizing genes that induce flat morphology (or ''flat reversion'') when expressed in a v-Ki-ras-transformed NIH 3T3 cell line, DT (3). The Krev-1 gene (4), also known as rap1A, which encodes a Ras-related protein containing a region identical to the effector domain of Ras, was isolated in a previous study (5) by using a plasmid-based human fibroblast cDNA expression library. Using a similar approach, Cutler et al. (6) isolated another transformation suppressor gene, rsp-1, encoding a leucinerich-repeat protein. Recently, we performed a similar screen of a human fibroblast cDNA expression library constructed with a new phagemid shuttle vector and obtained two cDNA clones exhibiting significant biological activities. One of these, clone CT124, was found to encode a truncated form of the MSX-2 homeobox protein, which induces flat reversion through a dominant-negative mechanism over the endogenous MSX-2 protein (7).Here we describe some properties of the other reversioninducing gene named RECK (reversion-inducing-cysteine-rich protein with Kazal motifs) and its product. This reversioninducing gene is unique in that it encodes an extracellular protein with protease inhibitor-like domains and its expression is suppressed strongly in many tumors and cells transformed by various kinds of oncogenes. Restored expression of the RECK gene inhibits the invasive and metastatic activities of tumor cells. We also found that RECK negatively regulates matrix metalloproteinase-9 (MMP-9) (8) in two ways: suppression of MMP-9 secretion from the cells and direct inhibition of its enzymatic activity. T...

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“…Each strain was grown on a plate containing 2% skim milk, 1% lactose, and 0.1% yeast extract at 37ЊC for 24 h. The temperature was then shifted to 25ЊC, and incubation was continued for 120 h. The assay plate was not supplemented [SSI(Ϫ)] or was supplemented [1 g/ml of medium; SSI(ϩ)] with SSI. (11,40) have demonstrated that the metalloproteases SGMPI and SGMPII, isolated from pronase P of S. griseus, are strongly inhibited by serine protease inhibitors such as SSI, alkaline protease inhibitor (API-2cЈ) (29), and plasminostreptin (12) as well as a metalloprotease inhibitor (SMPI from Streptomyces nigrescens TK-24) (20). Therefore, grouping of proteases and their inhibitors on the basis of their individual genus or species specificities should also be considered from the viewpoint of their physiological functions.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of a gene encoding extracellular serine protease isolated from a subtilisin inhibitor-deficient mutant of Streptomyces albogriseolus S-3253

Taguchi

Odaka

Watanabe

et al. 1995

Appl Environ Microbiol

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An extracellular serine protease produced by a mutant, M1, derived from Streptomyces albogriseolus S-3253 that no longer produces a protease inhibitor (Streptomyces subtilisin inhibitor [SSI]) was isolated. A 20-kDa protein was purified by its affinity for SSI and designated SAM-P20. The amino acid sequence of the aminoterminal region of SAM-P20 revealed high homology with the sequences of Streptomyces griseus proteases A and B, and the gene sequence confirmed the relationships. The sequence also revealed a putative amino acid signal sequence for SAM-P20 that apparently functioned to allow secretion of SAM-P20 from Escherichia coli carrying the recombinant gene. SAM-P20 produced by E. coli cells was shown to be sensitive to SSI inhibition.

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Interactions of Streptomyces Serine-Protease Inhibitors with Streptomyces griseus Metalloendopeptidase II

Cited by 16 publications

References 0 publications

The N‐terminal peptide of the transglutaminase‐activating metalloprotease inhibitor from Streptomyces mobaraensis accommodates both inhibition and glutamine cross‐linking sites

The N‐terminal peptide of the transglutaminase‐activating metalloprotease inhibitor from Streptomyces mobaraensis accommodates both inhibition and glutamine cross‐linking sites

Regulation of matrix metalloproteinase-9 and inhibition of tumor invasion by the membrane-anchored glycoprotein RECK

Molecular characterization of a gene encoding extracellular serine protease isolated from a subtilisin inhibitor-deficient mutant of Streptomyces albogriseolus S-3253

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