Detail of GM0001


Summary

The genetically modified enzybiotic, named ClyS , constructed by Domains Assembly strategy, change or extend lytic spectrum.
With  ClyS
  Sequence Length:  280 AA.
  Mass:  31998 Da.
  Isoelectric Point:  9.76
  Function:  ClyS lysed MRSA, vancomycin-intermediate strains of S. aureus (VISA), and methicillin-sensitive (MSSA) strains of S. aureus in vitro. In a mouse nasal decolonization model, a 2-log reduction in the viability of MRSA cells was seen 1 h following a single treatment with ClyS. One intraperitoneal dose of ClyS also protected against death by MRSA in a mouse septicemia model.

Construction

GM0001, constructed by Domains Assembly strategy.
    No schema construted on GM0001
Details:
GM0001[1-186]: derive from protein O56788 sequence 1-186

O56788, 467 AA., the N-acetylmuramoyl-L-alanine amidase from Staphylococcus phage Twort

Source: Staphylococcus phage Twort

Domains and repeats

4-141:  IPR007921, the CHAP domain
172-343:  IPR002502, the N-acetylmuramoyl-L-alanine amidase domain
355-382:  IPR003646, the SH3-like domain, bacterial-type

GO term prediction

Biological Process:  0009253, the peptidoglycan catabolic process
Molecular Function:  0008745, the N-acetylmuramoyl-L-alanine amidase activity


Linking to UniprotKB
Linging to InterPro
GM0001[187-280]: derive from protein A0EX11 sequence 158-251

A0EX11, 251 AA., the Amidase from Staphylococcus phage phiNM3

Source: Staphylococcus phage phiNM3

Domains and repeats

4-133:  IPR007921, the CHAP domain

GO term prediction

    No GO info found on GM0001

Linking to UniprotKB
Linging to InterPro

Annotation

1-186
187-280

1-186
: EAD, CHAP domain, derived from S. aureus Twort phage lysin;
187-280
: CBD, Cell wall-targeting domain, derived from S. aureus phage lysin (phiNM3);
Domain and repeats:
4-141
: IPR007921,CHAP domain;
GO term prediction:
MKTLKQAESYIKSKVNTGTDFDGLYGYQCMDLAVDYIYHVTDGKIRMWGNAKDAINNSFGGTATVYKNYPAFRPKYGDVVVWTTGNFATYGHIAIVTNPDPYGDLQYVTVLEQNWNGNGIYKTELATIRTHDYTGITHFIRPNFATESSVKKKDTKKKPKPSNRDGINKDKIVYDRTNINYNMVKR-GKSASKITVGSKAPYNLKWSKGAYFNAKIDGLGATSATRYGDNRTNYRFDVGQAVYAPGTLIYVFEIIDGWCRIYWNNHNEWIWHERLIVKEVF
  BlastP to GMEnzy

Production

  1.  Expressed by pJML6 in E. coli DH5Alpha,  Purified by CM-Sepharose column

Activity

  1.  In vitro activity of ClyS test on Staphylococcus aureus (S. aureus 8325-4 cells) by the turbidity-reduction assays
  showed the OD600 dropped 3- to 4-fold within ~15 min

Reference

  1. Anu Daniel,Chad Euler,Mattias Collin,Peter Chahales,Kenneth J. Gorelick, and Vincent A. Fischetti. (2010) Synergism between a Novel Chimeric Lysin and Oxacillin Protects against Infection by Methicillin-Resistant Staphylococcus aureus. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY. 544:1603-1613. [doi:10.1128/AAC.01625-09] [PMID:20086153] [FULL TEXT]
  2. Pastagia, M.Euler, C.Chahales, P.Fuentes-Duculan, J.Krueger, J. G.Fischetti, V. A.. (2011) A novel chimeric lysin shows superiority to mupirocin for skin decolonization of methicillin-resistant and -sensitive Staphylococcus aureus strains. Antimicrobial agents and chemotherapy. 02:738-744. [doi:10.1128/AAC.00890-10] [PMID:21098252] [FULL TEXT]

Comments

  •   [1]  Staphylococcus aureus is a major human pathogen responsible for a number of serious and sometimes fatal infections. One of its reservoirs on the human body is the skin, which is known to be a source of invasive infection. The potential for an engineered staphylococcus-specific phage lysin (ClyS) to be used for topical decolonization is presented. We formulated ClyS into an ointment and applied it to a mouse model of skin colonization/infection with S. aureus. Unlike the standard topical antibacterial agent mupirocin, ClyS eradicated a significantly greater number of methicillin-susceptible S. aureus (MSSA) and -resistant S. aureus (MRSA) bacteria: a 3-log reduction with ClyS as opposed to a 2-log reduction with mupirocin in our model. The use of ClyS also demonstrated a decreased potential for the development of resistance by MRSA and MSSA organisms compared to that from the use of mupirocin in vitro. Because antibodies may affect enzyme function, we tested ntibodies developed after repeated ClyS exposure for their effect on ClyS killing ability. Our results showed no inhibition of ClyS activity at various antibody titers. These data demonstrate the potential of developing ClyS as a novel class of topical antimicrobial agents specific to staphylococcus. See Antimicrobial agents and chemotherapy. 02:738-744 in detail.
  •   ----  ak0526@163.com   at  2013-09-05 16:16:50


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