Unidirectional binding of clostridial collagenase to triple helical substrates

Sagaya Theresa Leena Philominathan, Takaki Koide, Kentaro Hamada, Hiroyuki Yasui, Soenke Seifert, Osamu Matsushita, Joshua Sakon

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Histotoxic clostridia produce collagenases responsible for extensive tissue destruction in gas gangrene. The C-terminal collagen-binding domain (CBD) of these enzymes is the minimal segment required to bind to collagen fibril. Collagen binding efficiency of CBD is more pronounced in the presence of Ca2+. We have shown that CBD can be functional to anchor growth factors in local tissue. A 1H-15N HSQC NMR titration study with three different tropocollagen analogues ((POG)10)3, ((GPOG)7PRG)3, and (GPRG(POG)7C-carbamidomethyl)3, mapped a saddle-like binding cleft on CBD. NMR titrations with three nitroxide spin-labeled analogues of collagenous peptide, (PROXYL-G(POG)7PRG)3, (PROXYL-G(POG)7)3, and (GPRG(POG)7C-PROXYL)3 (where PROXYL represents 2,2,5,5-tetramethyl-L-pyrrolidinyloxy), unambiguously demonstrated unidirectional binding of CBD to the tropocollagen analogues. Small angle x-ray scattering data revealed that CBD binds closer to a terminus for each of the five different tropocollagen analogues, which in conjunction with NMR titration studies, implies a binding mode where CBD binds to the C terminus of the triple helix.

Original languageEnglish
Pages (from-to)10868-10876
Number of pages9
JournalJournal of Biological Chemistry
Volume284
Issue number16
DOIs
Publication statusPublished - Apr 17 2009
Externally publishedYes

Fingerprint

Collagenases
Collagen
Tropocollagen
Substrates
Titration
Nuclear magnetic resonance
Gas Gangrene
Tissue
Clostridium
Anchors
Intercellular Signaling Peptides and Proteins
Gases
X-Rays
Scattering
X rays
Peptides
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Philominathan, S. T. L., Koide, T., Hamada, K., Yasui, H., Seifert, S., Matsushita, O., & Sakon, J. (2009). Unidirectional binding of clostridial collagenase to triple helical substrates. Journal of Biological Chemistry, 284(16), 10868-10876. https://doi.org/10.1074/jbc.M807684200

Unidirectional binding of clostridial collagenase to triple helical substrates. / Philominathan, Sagaya Theresa Leena; Koide, Takaki; Hamada, Kentaro; Yasui, Hiroyuki; Seifert, Soenke; Matsushita, Osamu; Sakon, Joshua.

In: Journal of Biological Chemistry, Vol. 284, No. 16, 17.04.2009, p. 10868-10876.

Research output: Contribution to journalArticle

Philominathan, STL, Koide, T, Hamada, K, Yasui, H, Seifert, S, Matsushita, O & Sakon, J 2009, 'Unidirectional binding of clostridial collagenase to triple helical substrates', Journal of Biological Chemistry, vol. 284, no. 16, pp. 10868-10876. https://doi.org/10.1074/jbc.M807684200
Philominathan, Sagaya Theresa Leena ; Koide, Takaki ; Hamada, Kentaro ; Yasui, Hiroyuki ; Seifert, Soenke ; Matsushita, Osamu ; Sakon, Joshua. / Unidirectional binding of clostridial collagenase to triple helical substrates. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 16. pp. 10868-10876.
@article{be166335d1234881adb8d7b4a4f00f89,
title = "Unidirectional binding of clostridial collagenase to triple helical substrates",
abstract = "Histotoxic clostridia produce collagenases responsible for extensive tissue destruction in gas gangrene. The C-terminal collagen-binding domain (CBD) of these enzymes is the minimal segment required to bind to collagen fibril. Collagen binding efficiency of CBD is more pronounced in the presence of Ca2+. We have shown that CBD can be functional to anchor growth factors in local tissue. A 1H-15N HSQC NMR titration study with three different tropocollagen analogues ((POG)10)3, ((GPOG)7PRG)3, and (GPRG(POG)7C-carbamidomethyl)3, mapped a saddle-like binding cleft on CBD. NMR titrations with three nitroxide spin-labeled analogues of collagenous peptide, (PROXYL-G(POG)7PRG)3, (PROXYL-G(POG)7)3, and (GPRG(POG)7C-PROXYL)3 (where PROXYL represents 2,2,5,5-tetramethyl-L-pyrrolidinyloxy), unambiguously demonstrated unidirectional binding of CBD to the tropocollagen analogues. Small angle x-ray scattering data revealed that CBD binds closer to a terminus for each of the five different tropocollagen analogues, which in conjunction with NMR titration studies, implies a binding mode where CBD binds to the C terminus of the triple helix.",
author = "Philominathan, {Sagaya Theresa Leena} and Takaki Koide and Kentaro Hamada and Hiroyuki Yasui and Soenke Seifert and Osamu Matsushita and Joshua Sakon",
year = "2009",
month = "4",
day = "17",
doi = "10.1074/jbc.M807684200",
language = "English",
volume = "284",
pages = "10868--10876",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "16",

}

TY - JOUR

T1 - Unidirectional binding of clostridial collagenase to triple helical substrates

AU - Philominathan, Sagaya Theresa Leena

AU - Koide, Takaki

AU - Hamada, Kentaro

AU - Yasui, Hiroyuki

AU - Seifert, Soenke

AU - Matsushita, Osamu

AU - Sakon, Joshua

PY - 2009/4/17

Y1 - 2009/4/17

N2 - Histotoxic clostridia produce collagenases responsible for extensive tissue destruction in gas gangrene. The C-terminal collagen-binding domain (CBD) of these enzymes is the minimal segment required to bind to collagen fibril. Collagen binding efficiency of CBD is more pronounced in the presence of Ca2+. We have shown that CBD can be functional to anchor growth factors in local tissue. A 1H-15N HSQC NMR titration study with three different tropocollagen analogues ((POG)10)3, ((GPOG)7PRG)3, and (GPRG(POG)7C-carbamidomethyl)3, mapped a saddle-like binding cleft on CBD. NMR titrations with three nitroxide spin-labeled analogues of collagenous peptide, (PROXYL-G(POG)7PRG)3, (PROXYL-G(POG)7)3, and (GPRG(POG)7C-PROXYL)3 (where PROXYL represents 2,2,5,5-tetramethyl-L-pyrrolidinyloxy), unambiguously demonstrated unidirectional binding of CBD to the tropocollagen analogues. Small angle x-ray scattering data revealed that CBD binds closer to a terminus for each of the five different tropocollagen analogues, which in conjunction with NMR titration studies, implies a binding mode where CBD binds to the C terminus of the triple helix.

AB - Histotoxic clostridia produce collagenases responsible for extensive tissue destruction in gas gangrene. The C-terminal collagen-binding domain (CBD) of these enzymes is the minimal segment required to bind to collagen fibril. Collagen binding efficiency of CBD is more pronounced in the presence of Ca2+. We have shown that CBD can be functional to anchor growth factors in local tissue. A 1H-15N HSQC NMR titration study with three different tropocollagen analogues ((POG)10)3, ((GPOG)7PRG)3, and (GPRG(POG)7C-carbamidomethyl)3, mapped a saddle-like binding cleft on CBD. NMR titrations with three nitroxide spin-labeled analogues of collagenous peptide, (PROXYL-G(POG)7PRG)3, (PROXYL-G(POG)7)3, and (GPRG(POG)7C-PROXYL)3 (where PROXYL represents 2,2,5,5-tetramethyl-L-pyrrolidinyloxy), unambiguously demonstrated unidirectional binding of CBD to the tropocollagen analogues. Small angle x-ray scattering data revealed that CBD binds closer to a terminus for each of the five different tropocollagen analogues, which in conjunction with NMR titration studies, implies a binding mode where CBD binds to the C terminus of the triple helix.

UR - http://www.scopus.com/inward/record.url?scp=67449099703&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67449099703&partnerID=8YFLogxK

U2 - 10.1074/jbc.M807684200

DO - 10.1074/jbc.M807684200

M3 - Article

VL - 284

SP - 10868

EP - 10876

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 16

ER -