Design and synthesis of sensitive fluorogenic substrates specific for Lys-gingipain

Naoko Abe, Atsuyo Baba, Tomoko Kadowaki, Kuniaki Okamoto, Shinji Okazaki, Tetsuji Asao, Kenji Yamamoto

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Lys-gingipain (Kgp) is a major cysteine proteinase produced by the oral anaerobic bacterium Porphyromonas gingivalis, and has been implicated as a major pathogen in the development and progression of advanced adult periodontitis. This enzyme is believed to act as a major virulence factor of the disease, yet there exist no convenient and sensitive substrates for analyzing its biological activity. For a better understanding of the importance of this enzyme in the organism, there is an urgent need for specific substrates. Here we designed and synthesized two peptide 4-methyl-coumaryl-7-amides (MCA), carbobenzoxy (Z)-His-Glu-Lys-MCA, and Z-Glu-Lys-MCA, and tested their possible use as sensitive substrates for Kgp with limited specificity. Both substrates exhibited greater k(cat)/K(m) values than the best known Kgp substrates described so far. Both substrates were resistant to Arg-gingipain, another pathogenic cysteine proteinase from P. gingivalis, as well as trypsin and cathepsins B, L, and H. The levels of Kgp in various microorganisms and human cells were determined with Z-His-Glu-Lys-MCA. Little or no Kgp-like activity was detected in either other microorganisms or human cells tested. These results indicate that the present substrates are a valuable and fast tool for routine assays and for mechanistic studies on Kgp.

Original languageEnglish
Pages (from-to)877-881
Number of pages5
JournalJournal of Biochemistry
Volume128
Issue number5
Publication statusPublished - 2000
Externally publishedYes

Fingerprint

Fluorescent Dyes
Cysteine Proteases
Substrates
Cathepsin H
Cathepsin L
Chronic Periodontitis
Cathepsin B
Porphyromonas gingivalis
Anaerobic Bacteria
Microorganisms
Virulence Factors
Enzymes
Trypsin
Cells
Peptides
Pathogens
Bioactivity
Lys-gingipain
4-methyl-coumaryl-7-amide
Assays

Keywords

  • Cysteine proteinase
  • Fluorogenic substrate
  • Lys-gingipain
  • Periodontitis
  • Porphyromonas gingivalis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Abe, N., Baba, A., Kadowaki, T., Okamoto, K., Okazaki, S., Asao, T., & Yamamoto, K. (2000). Design and synthesis of sensitive fluorogenic substrates specific for Lys-gingipain. Journal of Biochemistry, 128(5), 877-881.

Design and synthesis of sensitive fluorogenic substrates specific for Lys-gingipain. / Abe, Naoko; Baba, Atsuyo; Kadowaki, Tomoko; Okamoto, Kuniaki; Okazaki, Shinji; Asao, Tetsuji; Yamamoto, Kenji.

In: Journal of Biochemistry, Vol. 128, No. 5, 2000, p. 877-881.

Research output: Contribution to journalArticle

Abe, N, Baba, A, Kadowaki, T, Okamoto, K, Okazaki, S, Asao, T & Yamamoto, K 2000, 'Design and synthesis of sensitive fluorogenic substrates specific for Lys-gingipain', Journal of Biochemistry, vol. 128, no. 5, pp. 877-881.
Abe, Naoko ; Baba, Atsuyo ; Kadowaki, Tomoko ; Okamoto, Kuniaki ; Okazaki, Shinji ; Asao, Tetsuji ; Yamamoto, Kenji. / Design and synthesis of sensitive fluorogenic substrates specific for Lys-gingipain. In: Journal of Biochemistry. 2000 ; Vol. 128, No. 5. pp. 877-881.
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AU - Asao, Tetsuji

AU - Yamamoto, Kenji

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