Update 1 of: Proteases Universally Recognize Beta Strands In Their Active Sites

Madala, Praveen K.; Tyndall, Joel D. A.; Nall, Tessa; Fairlie, David P.

doi:10.1021/cr900368a

Cited by 152 publications

(150 citation statements)

References 221 publications

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“…Furthermore, a proper processing requires cleavage sites located in close proximity to the central hydrophobic and carboxyl-terminal hydrophilic border (reviewed in reference 41). Moreover, those proteins processed by proteases tend to be cleaved between elements of secondary structure rather than within helical, turn, or sheet regions (26). In this sense, the secondarystructure prediction of the native AS-48 preprotein using the PHD algorithm (http://www.predictprotein.org/) suggests that the SP (35 residues) has a propensity to form an ␣-helical conformation (positions 13 to 31) in wild-type AS-48.…”

Section: Resultsmentioning

confidence: 99%

Insights into the Functionality of the Putative Residues Involved in Enterocin AS-48 Maturation

Cebrián

Maqueda

Neira

et al. 2010

Appl Environ Microbiol

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AS-48 is a 70-residue, ␣-helical, cationic bacteriocin produced by Enterococcus faecalis and is very singular in its circular structure and its broad antibacterial spectrum. The AS-48 preprotein consists of an N-terminal signal peptide (SP) (35 residues) followed by a proprotein moiety that undergoes posttranslational modifications to yield the mature and active circular protein. For the study of the specificity of the region of AS-48 that is responsible for maturation, three single mutants have been generated by site-directed mutagenesis in the as-48A structural gene. The substitutions were made just in the residues that are thought to constitute a recognition site for the SP cleavage enzyme (His-1, Met1) and in those involved in circularization (Met1, Trp70). Each derivative was expressed in the enterococcal JH2-2 strain containing the necessary native biosynthetic machinery for enterocin production. The importance of these derivatives in AS-48 processing has been evaluated on the basis of the production and structural characterization of the corresponding derivatives. Notably, only two of them (Trp70Ala and Met1Ala derivatives) could be purified in different forms and amounts and are characterized for their bactericidal activity and secondary structure. We could not detect any production of AS-48 in JH2-2(pAM401-81 His-1Ile ) by using the conventional chromatographic techniques, despite the high efficiency of the culture conditions applied to produce this enterocin. Our results underline the different important roles of the mutated residues in (i) the elimination of the SP, (ii) the production levels and antibacterial activity of the mature proteins, and (iii) protein circularization. Moreover, our findings suggest that His-1 is critically involved in cleavage site recognition, its substitution being responsible for the blockage of processing, thereby hampering the production of the specific protein in the cellular culture supernatant.

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Section: Resultsmentioning

confidence: 99%

Insights into the Functionality of the Putative Residues Involved in Enterocin AS-48 Maturation

Cebrián

Maqueda

Neira

et al. 2010

Appl Environ Microbiol

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“…For infectious organisms such as bacteria, viruses and parasites, generally need protease inhibitors and HIV has been successfully targeted by protease inhibitors and these are used clinically (Amprenavir, Atazanavir, etc.) 20 .…”

Section: Discussionmentioning

confidence: 99%

“…The search for new -lactamase inhibitors, thus, has a remarkable clinical and industrial potential. In this work, enzyme inhibition studies of the crude extracts were investigated with above mentioned enzymes in search for potential substrates with inhibitory nature 18,19,20 . In our previous study, we have reported the antioxidant activity of H. lupulus extracts with solvents including n-hexane, acetone, methanol (1, 2 and 3) and 25% aqueous ethanol.…”

Section: Introductionmentioning

confidence: 99%

Antiproliferative activity ofHumulus lupulusextracts on human hepatoma (Hep3B), colon (HT-29) cancer cells and proteases, tyrosinase,β-lactamase enzyme inhibition studies

Önder

Türkoğlu

et al. 2015

Journal of Enzyme Inhibition and Medicinal Chemistry

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The aims of this study were to examine the antiproliferation of Humulus lupulus extracts on human hepatoma carcinoma (Hep3B) and human colon carcinoma (HT-29) cell lines along with enzyme inhibitory effects of the crude extracts. Potential cell cytotoxicity of six different H. lupulus extracts were assayed on various cancer cells using MTT assay at 24, 48 and 72 h intervals. Methanol-1 extract has inhibited the cell proliferation with doses of 0.6-1 mg/mL in a time dependent (48 and 72 hours) manner in Hep3B cells with 70% inhibition, while inhibitory effect was not seen in colon cancer cells. Acetone extract has increased the cell proliferation at low doses of 0.1 mg/mL for 72 h in Hep3B cells and 0.1-0.2 mg/mL for 48 and 72 h in HT29 cells. The inhibitory effects of the extracts were compared by relative maximum activity values (V max ) using proteases such as -chymotrypsin, trypsin and papain, tyrosinase and -lactamase (penicillinase).

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“…The edge strand of zincins in general plays a critical role in substrate binding through the formation of a ladder of mainchain hydrogen bonds with the substrate (37). The edge strand is the only antiparallel strand of the N-terminal ␤-sheet.…”

Section: Discussionmentioning

confidence: 99%

The Structure of Mlc Titration Factor A (MtfA/YeeI) Reveals a Prototypical Zinc Metallopeptidase Related to Anthrax Lethal Factor

Göhler

Kosfeld

et al. 2012

J Bacteriol

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MtfA of Escherichia coli (formerly YeeI) was previously identified as a regulator of the phosphoenolpyruvate (PEP)-dependent:glucose phosphotransferase system. MtfA homolog proteins are highly conserved, especially among beta- and gammaproteobacteria. We determined the crystal structures of the full-length MtfA apoenzyme from Klebsiella pneumoniae and its complex with zinc (holoenzyme) at 2.2 and 1.95 Å, respectively. MtfA contains a conserved H 149 E 150 XXH 153 +E 212 +Y 205 metallopeptidase motif. The presence of zinc in the active site induces significant conformational changes in the region around Tyr205 compared to the conformation of the apoenzyme. Additionally, the zinc-bound MtfA structure is in a self-inhibitory conformation where a region that was disordered in the unliganded structure is now observed in the active site and a nonproductive state of the enzyme is formed. MtfA is related to the catalytic domain of the anthrax lethal factor and the Mop protein involved in the virulence of Vibrio cholerae , with conservation in both overall structure and in the residues around the active site. These results clearly provide support for MtfA as a prototypical zinc metallopeptidase (gluzincin clan).

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Update 1 of: Proteases Universally Recognize Beta Strands In Their Active Sites

Cited by 152 publications

References 221 publications

Insights into the Functionality of the Putative Residues Involved in Enterocin AS-48 Maturation

Insights into the Functionality of the Putative Residues Involved in Enterocin AS-48 Maturation

Antiproliferative activity ofHumulus lupulusextracts on human hepatoma (Hep3B), colon (HT-29) cancer cells and proteases, tyrosinase,β-lactamase enzyme inhibition studies

The Structure of Mlc Titration Factor A (MtfA/YeeI) Reveals a Prototypical Zinc Metallopeptidase Related to Anthrax Lethal Factor

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