Involvement of the Arg566 residue of Aeromonas sobria serine protease in substrate specificity

Hidetomo Kobayashi, Tadamune Otsubo, Fumiteru Teraoka, Kiyoshi Ikeda, Soshi Seike, Eizo Takahashi, Keinosuke Okamoto, Toru Yoshida, Hideaki Tsuge, Hiroyasu Yamanaka

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Aeromonas sobria serine protease (ASP) is an extracellular serine protease secreted by the organism. Here, we identified the amino acid residue of ASP that contributes to substrate specificity by using both synthetic peptides and biological protein components. The results showed that the arginine residue at position 566 (Arg-566) of ASP, which is located in the extra occluding region of ASP close to an entrance of the catalytic cavity, is involved in the substrate specificity. A substitutional point mutation of the Arg-566 residue of ASP to Ala residue (ASP[R566A]) caused a decrease of the proteolytic efficiency for a certain substrate. In addition, ASP lost the ability to recognize the primary substrate by such a point mutation, and ASP[R566A] reacted to a wide range of synthetic substrates. It is likely that Arg-566 causes an interaction with the amino acid residue at position P3 of the substrate, which is the third amino acid residue upstream from the cleavage site. Another study using ORF2 protein, a chaperone protein of ASP, further suggested that Arg-566 could also play an important role in interaction with ORF2. We therefore conclude that the Arg-566 residue of ASP is likely responsible for the selection of substrates.

Original languageEnglish
Article numbere0186392
JournalPLoS One
Volume12
Issue number10
DOIs
Publication statusPublished - Oct 1 2017
Externally publishedYes

Fingerprint

Aeromonas sobria
Aeromonas
Serine Proteases
serine proteinases
substrate specificity
Substrate Specificity
Substrates
arginine
Arginine
point mutation
Point Mutation
Amino Acids
amino acids
Proteins
proteins
synthetic peptides

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Kobayashi, H., Otsubo, T., Teraoka, F., Ikeda, K., Seike, S., Takahashi, E., ... Yamanaka, H. (2017). Involvement of the Arg566 residue of Aeromonas sobria serine protease in substrate specificity. PLoS One, 12(10), [e0186392]. https://doi.org/10.1371/journal.pone.0186392

Involvement of the Arg566 residue of Aeromonas sobria serine protease in substrate specificity. / Kobayashi, Hidetomo; Otsubo, Tadamune; Teraoka, Fumiteru; Ikeda, Kiyoshi; Seike, Soshi; Takahashi, Eizo; Okamoto, Keinosuke; Yoshida, Toru; Tsuge, Hideaki; Yamanaka, Hiroyasu.

In: PLoS One, Vol. 12, No. 10, e0186392, 01.10.2017.

Research output: Contribution to journalArticle

Kobayashi, H, Otsubo, T, Teraoka, F, Ikeda, K, Seike, S, Takahashi, E, Okamoto, K, Yoshida, T, Tsuge, H & Yamanaka, H 2017, 'Involvement of the Arg566 residue of Aeromonas sobria serine protease in substrate specificity', PLoS One, vol. 12, no. 10, e0186392. https://doi.org/10.1371/journal.pone.0186392
Kobayashi, Hidetomo ; Otsubo, Tadamune ; Teraoka, Fumiteru ; Ikeda, Kiyoshi ; Seike, Soshi ; Takahashi, Eizo ; Okamoto, Keinosuke ; Yoshida, Toru ; Tsuge, Hideaki ; Yamanaka, Hiroyasu. / Involvement of the Arg566 residue of Aeromonas sobria serine protease in substrate specificity. In: PLoS One. 2017 ; Vol. 12, No. 10.
@article{492c8d6908c14c48aaf2319b915bdff2,
title = "Involvement of the Arg566 residue of Aeromonas sobria serine protease in substrate specificity",
abstract = "Aeromonas sobria serine protease (ASP) is an extracellular serine protease secreted by the organism. Here, we identified the amino acid residue of ASP that contributes to substrate specificity by using both synthetic peptides and biological protein components. The results showed that the arginine residue at position 566 (Arg-566) of ASP, which is located in the extra occluding region of ASP close to an entrance of the catalytic cavity, is involved in the substrate specificity. A substitutional point mutation of the Arg-566 residue of ASP to Ala residue (ASP[R566A]) caused a decrease of the proteolytic efficiency for a certain substrate. In addition, ASP lost the ability to recognize the primary substrate by such a point mutation, and ASP[R566A] reacted to a wide range of synthetic substrates. It is likely that Arg-566 causes an interaction with the amino acid residue at position P3 of the substrate, which is the third amino acid residue upstream from the cleavage site. Another study using ORF2 protein, a chaperone protein of ASP, further suggested that Arg-566 could also play an important role in interaction with ORF2. We therefore conclude that the Arg-566 residue of ASP is likely responsible for the selection of substrates.",
author = "Hidetomo Kobayashi and Tadamune Otsubo and Fumiteru Teraoka and Kiyoshi Ikeda and Soshi Seike and Eizo Takahashi and Keinosuke Okamoto and Toru Yoshida and Hideaki Tsuge and Hiroyasu Yamanaka",
year = "2017",
month = "10",
day = "1",
doi = "10.1371/journal.pone.0186392",
language = "English",
volume = "12",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "10",

}

TY - JOUR

T1 - Involvement of the Arg566 residue of Aeromonas sobria serine protease in substrate specificity

AU - Kobayashi, Hidetomo

AU - Otsubo, Tadamune

AU - Teraoka, Fumiteru

AU - Ikeda, Kiyoshi

AU - Seike, Soshi

AU - Takahashi, Eizo

AU - Okamoto, Keinosuke

AU - Yoshida, Toru

AU - Tsuge, Hideaki

AU - Yamanaka, Hiroyasu

PY - 2017/10/1

Y1 - 2017/10/1

N2 - Aeromonas sobria serine protease (ASP) is an extracellular serine protease secreted by the organism. Here, we identified the amino acid residue of ASP that contributes to substrate specificity by using both synthetic peptides and biological protein components. The results showed that the arginine residue at position 566 (Arg-566) of ASP, which is located in the extra occluding region of ASP close to an entrance of the catalytic cavity, is involved in the substrate specificity. A substitutional point mutation of the Arg-566 residue of ASP to Ala residue (ASP[R566A]) caused a decrease of the proteolytic efficiency for a certain substrate. In addition, ASP lost the ability to recognize the primary substrate by such a point mutation, and ASP[R566A] reacted to a wide range of synthetic substrates. It is likely that Arg-566 causes an interaction with the amino acid residue at position P3 of the substrate, which is the third amino acid residue upstream from the cleavage site. Another study using ORF2 protein, a chaperone protein of ASP, further suggested that Arg-566 could also play an important role in interaction with ORF2. We therefore conclude that the Arg-566 residue of ASP is likely responsible for the selection of substrates.

AB - Aeromonas sobria serine protease (ASP) is an extracellular serine protease secreted by the organism. Here, we identified the amino acid residue of ASP that contributes to substrate specificity by using both synthetic peptides and biological protein components. The results showed that the arginine residue at position 566 (Arg-566) of ASP, which is located in the extra occluding region of ASP close to an entrance of the catalytic cavity, is involved in the substrate specificity. A substitutional point mutation of the Arg-566 residue of ASP to Ala residue (ASP[R566A]) caused a decrease of the proteolytic efficiency for a certain substrate. In addition, ASP lost the ability to recognize the primary substrate by such a point mutation, and ASP[R566A] reacted to a wide range of synthetic substrates. It is likely that Arg-566 causes an interaction with the amino acid residue at position P3 of the substrate, which is the third amino acid residue upstream from the cleavage site. Another study using ORF2 protein, a chaperone protein of ASP, further suggested that Arg-566 could also play an important role in interaction with ORF2. We therefore conclude that the Arg-566 residue of ASP is likely responsible for the selection of substrates.

UR - http://www.scopus.com/inward/record.url?scp=85031121176&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85031121176&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0186392

DO - 10.1371/journal.pone.0186392

M3 - Article

C2 - 29023605

AN - SCOPUS:85031121176

VL - 12

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 10

M1 - e0186392

ER -