title: Recombinant fragilysin isoforms cause E-cadherin cleavage of intact cells and do not cleave isolated E-cadherin.
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.
authors.affiliation: 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.
authors.forename: Daria
authors.initials: D
authors.lastname: Kharlampieva
authors.affiliation: 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.
authors.forename: Valentin
authors.initials: V
authors.lastname: Manuvera
authors.affiliation: 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.
authors.forename: Oleg
authors.initials: O
authors.lastname: Podgorny
authors.affiliation: 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.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Grafskaia
authors.affiliation: 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.
authors.forename: Sergey
authors.initials: S
authors.lastname: Kovalchuk
authors.affiliation: Scientific Research Institute of Physical-Chemical Medicine of the Federal Medical and Biological Agency, Malaya Pirogovskaya str. 1a, Moscow 119435, Russia.
authors.forename: Olga
authors.initials: O
authors.lastname: Pobeguts
authors.affiliation: 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.
authors.forename: Ilya
authors.initials: I
authors.lastname: Altukhov
authors.affiliation: 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.
authors.forename: Vadim
authors.initials: V
authors.lastname: Govorun
authors.affiliation: 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.
authors.forename: Vassili
authors.initials: V
authors.lastname: Lazarev
mesh_terms: D001441:Bacteroides fragilis / Q000201:enzymology / Q000235:genetics; D015820:Cadherins / Q000378:metabolism*; D002460:Cell Line; D004847:Epithelial Cells / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D059748:Proteolysis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D015820:Cadherins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D008666:Metalloendopeptidases; C091198:fragilysin
keywords: BFT; Bacteroides fragilis; E-cadherin; Fragilysin; Metalloprotease