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

Daria Kharlampieva; Valentin Manuvera; Oleg Podgorny; Ekaterina Grafskaia; Sergey Kovalchuk; Olga Pobeguts; Ilya Altukhov; Vadim Govorun; Vassili Lazarev

doi:10.1016/j.micpath.2015.05.003

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

Microb Pathog. 2015 Jun-Jul:83-84:47-56. doi: 10.1016/j.micpath.2015.05.003. Epub 2015 May 18.

Authors

Daria Kharlampieva¹, Valentin Manuvera², Oleg Podgorny³, Ekaterina Grafskaia², Sergey Kovalchuk⁴, Olga Pobeguts⁵, Ilya Altukhov², Vadim Govorun⁶, Vassili Lazarev²

Affiliations

¹ Scientific Research Institute of Physical-Chemical Medicine of the Federal Medical and Biological Agency, Malaya Pirogovskaya str. 1a, Moscow 119435, Russia. Electronic address: harlampieva_d@mail.ru.
² Scientific Research Institute of Physical-Chemical Medicine of the Federal Medical and Biological Agency, Malaya Pirogovskaya str. 1a, Moscow 119435, Russia; Moscow Institute of Physics and Technology, Institutskiy per. 9, Dolgoprudny, Moscovskaya obl 141700, Russia.
³ Scientific Research Institute of Physical-Chemical Medicine of the Federal Medical and Biological Agency, Malaya Pirogovskaya str. 1a, Moscow 119435, Russia; Moscow Institute of Physics and Technology, Institutskiy per. 9, Dolgoprudny, Moscovskaya obl 141700, Russia; Koltzov Institute of Developmental Biology of the Russian Academy of Sciences, Vavilov str. 26, Moscow 119334, Russia.
⁴ Scientific Research Institute of Physical-Chemical Medicine of the Federal Medical and Biological Agency, Malaya Pirogovskaya str. 1a, Moscow 119435, Russia; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Miklukho-Maklaya str. 16/10, Moscow 117997, Russia.
⁵ Scientific Research Institute of Physical-Chemical Medicine of the Federal Medical and Biological Agency, Malaya Pirogovskaya str. 1a, Moscow 119435, Russia.
⁶ Scientific Research Institute of Physical-Chemical Medicine of the Federal Medical and Biological Agency, Malaya Pirogovskaya str. 1a, Moscow 119435, Russia; Moscow Institute of Physics and Technology, Institutskiy per. 9, Dolgoprudny, Moscovskaya obl 141700, Russia; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Miklukho-Maklaya str. 16/10, Moscow 117997, Russia.

PMID: 25998017
DOI: 10.1016/j.micpath.2015.05.003

Abstract

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 motif. At the same time recombinant BFTs did not cleave recombinant E-cadherin or E-cadherin in isolated cell fractions. It indicates that E-cadherin may be not direct substrate for BFT. We also detected and identified proteins released into the cultural medium after HT-29 cells treatment with BFT. The role of these proteins in pathogenesis and cell response to BFT remains to be determined.

Keywords: BFT; Bacteroides fragilis; E-cadherin; Fragilysin; Metalloprotease.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacteroides fragilis / enzymology
Bacteroides fragilis / genetics
Cadherins / metabolism*
Cell Line
Epithelial Cells / drug effects
Escherichia coli / genetics
Escherichia coli / metabolism
Humans
Metalloendopeptidases / genetics
Metalloendopeptidases / metabolism*
Protein Isoforms / genetics
Protein Isoforms / metabolism*
Proteolysis
Recombinant Proteins / genetics
Recombinant Proteins / metabolism

Substances

Cadherins
Protein Isoforms
Recombinant Proteins
Metalloendopeptidases
fragilysin