Proteomics

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HT-29 treated with mature fragilysin LC-MSMS

ABSTRACT: Fragilysin is a protein secreted by toxigenic Bacteroides fragilis strains. Fragilysin has a motive HEXXHXXGXXH, which is a zinc-binding motif found in metalloproteinases clan termed metzincins. In this study we obtained all three known recombinant fragilysin isoforms in Escherichia coli and tested its activity. We detected no cleavage of gelatin and chromogenic substrates (such as azocoll and azocasein) by all isoforms. We demonstrated that treatment of HT-29 cells with fragilysins caused E-cadherin cleavage, nevertheless cleavage of recombinant E-cadherins was not observe as well as E-cadherin in isolated cell fractions. It was shown that the native structure of active site characteristic for metalloproteinases is necessary for fragilysin biologic activity. We detected proteins that are released in cultural medium after HT-29 cells treatment with fragilysin. These proteins are potential substrates for fragilysin.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Colon

SUBMITTER: Ilya Altukhov  

LAB HEAD: Vasiliy Lazarev

PROVIDER: PXD001271 | Pride | 2015-05-26

REPOSITORIES: Pride

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Dataset's files

Source:
Action DRS
T302-20130412_HTK1_Rgest_500nlinj_IDA_1.wiff Wiff
T302-20130412_HTK1_Rgest_500nlinj_IDA_1.wiff.scan Wiff
T302-20130412_HTK1_Rgest_500nlinj_IDA_1_vs_swissprot_human_MGFPeaklist.mgf Mgf
T302_K_RG.dat Other
T302_K_RG.dat-pride.pride.mgf.gz Mgf
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Publications

Recombinant fragilysin isoforms cause E-cadherin cleavage of intact cells and do not cleave isolated E-cadherin.

Kharlampieva Daria D   Manuvera Valentin V   Podgorny Oleg O   Grafskaia Ekaterina E   Kovalchuk Sergey S   Pobeguts Olga O   Altukhov Ilya I   Govorun Vadim V   Lazarev Vassili V  

Microbial pathogenesis 20150518

The fragilysin (BFT) is a protein secreted by enterotoxigenic Bacteroides fragilis strains. BFT contains zinc-binding motif which was found in the metzincins family of metalloproteinases. In this study, we generated three known recombinant isoforms of BFT using Escherichia coli, tested their activity and examined whether E-cadherin is a substrate for BFTs. BFT treatment of HT-29 cells induced endogenous E-cadherin cleavage, and this BFT activity requires the native structure of zinc-binding moti  ...[more]

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