Enzymatic and Molecular Characteristics of the Efficiency and Specificity of Exfoliative Toxin Cleavage of Desmoglein 1

Yasushi Hanakawa, Norman M. Schechter, Chenyan Lin, Koji Nishifuji, Masayuki Amagai, John R. Stanley

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Exfoliative toxins (ETs) from Staphylococcus aureus blister the superficial epidermis by hydrolyzing a single peptide bond, Glu 381-Gly382, located between extracellular domains 3 and 4 of desmoglein 1 (Dsg1). Enzyme activity is dependent on the calcium-stabilized structure of Dsg1. Here we further define the characteristics of this cleavage. Kinetic studies monitoring the cleavage of Dsg1 by ETA, ETB, and ETD demonstrated kcat/Km values of 2-6 × 104 M-1, s-1 suggesting very efficient proteolysis. Proteolysis by ETA was not efficiently inhibited by broad spectrum serine protease inhibitors, suggesting that the enzyme cleavage site may be inactive or inaccessible before specific binding to its substrate. Using truncated mutants of human Dsg1 and chimeric molecules between human Dsg1 and either human Dsg3 or canine Dsg1, we show that for cleavage, human-specific amino acids from Dsg1 are necessary in extracellular domain 3 upstream of the scissile bond. If these residues are canine rather than human, ETA binds, but does not cleave, canine Dsg1. These data suggest that the exquisite specificity and efficiency of ETA may depend on the enzyme's binding upstream of the cleavage site with a very specific fit, like a key in a lock.

Original languageEnglish
Pages (from-to)5268-5277
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number7
DOIs
Publication statusPublished - 2004 Feb 13

Fingerprint

Exfoliatins
Desmoglein 1
Proteolysis
Canidae
Enzymes
Serine Proteinase Inhibitors
Enzyme activity
Blister
Epidermis
Staphylococcus aureus
Calcium
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Enzymatic and Molecular Characteristics of the Efficiency and Specificity of Exfoliative Toxin Cleavage of Desmoglein 1. / Hanakawa, Yasushi; Schechter, Norman M.; Lin, Chenyan; Nishifuji, Koji; Amagai, Masayuki; Stanley, John R.

In: Journal of Biological Chemistry, Vol. 279, No. 7, 13.02.2004, p. 5268-5277.

Research output: Contribution to journalArticle

Hanakawa, Yasushi ; Schechter, Norman M. ; Lin, Chenyan ; Nishifuji, Koji ; Amagai, Masayuki ; Stanley, John R. / Enzymatic and Molecular Characteristics of the Efficiency and Specificity of Exfoliative Toxin Cleavage of Desmoglein 1. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 7. pp. 5268-5277.
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