Enzymatic and molecular characteristics of the efficiency and specificity of exfoliative toxin cleavage of desmoglein 1

被引:41
作者
Hanakawa, Y
Schechter, NM
Lin, CY
Nishifuji, K
Amagai, M
Stanley, JR
机构
[1] Univ Penn, Dept Dermatol, Sch Med, Philadelphia, PA 19104 USA
[2] Keio Univ, Sch Med, Dept Dermatol, Tokyo 1608582, Japan
关键词
D O I
10.1074/jbc.M311087200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Exfoliative toxins (ETs) from Staphylococcus aureus blister the superficial epidermis by hydrolyzing a single peptide bond, Glu(381)-Gly(382), 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 k(cat)/K-m values of 2-6 x 10(4) 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.
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页码:5268 / 5277
页数:10
相关论文
共 39 条
[1]   Toxin in bullous impetigo and staphylococcal scalded-skin syndrome targets desmoglein 1 [J].
Amagai, M ;
Matsuyoshi, N ;
Wang, ZH ;
Andl, C ;
Stanley, JR .
NATURE MEDICINE, 2000, 6 (11) :1275-1277
[2]   EXTRACELLULAR DOMAIN OF PEMPHIGUS-VULGARIS ANTIGEN (DESMOGLEIN-3) MEDIATES WEAK HEMOPHILIC ADHESION [J].
AMAGAI, M ;
KARPATI, S ;
KLAUSKOVTUN, V ;
UDEY, MC ;
STANLEY, JR .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 1994, 102 (04) :402-408
[3]   Staphylococcal exfoliative toxin B specifically cleaves desmoglein 1 [J].
Amagai, M ;
Yamaguchi, T ;
Hanakawa, Y ;
Nishifuji, K ;
Sugai, M ;
Stanley, JR .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 2002, 118 (05) :845-850
[4]  
Angst BD, 2001, J CELL SCI, V114, P625
[5]  
[Anonymous], PROTEIN PROTEIN RECO
[6]   C-cadherin ectodomain structure and implications for cell adhesion mechanisms [J].
Boggon, TJ ;
Murray, J ;
Chappuis-Flament, S ;
Wong, E ;
Gumbiner, BM ;
Shapiro, L .
SCIENCE, 2002, 296 (5571) :1308-1313
[7]   The structure of Staphylococcus aureus epidermolytic toxin A, an atypic serine protease, at 1.7 angstrom resolution [J].
Cavarelli, J ;
Prevost, G ;
Bourguet, W ;
Moulinier, L ;
Chevrier, B ;
Delagoutte, B ;
Bilwes, A ;
Mourey, L ;
Rifai, S ;
Piemont, Y ;
Moras, D .
STRUCTURE, 1997, 5 (06) :813-824
[8]   Are desmosomes more than tethers for intermediate filaments? [J].
Green, KJ ;
Gaudry, CA .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2000, 1 (03) :208-216
[9]   Calcium-dependent conformation of desmoglein 1 is required for its cleavage by exfoliative toxin [J].
Hanakawa, Y ;
Selwood, T ;
Woo, D ;
Lin, C ;
Schechter, NM ;
Stanley, JR .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 2003, 121 (02) :383-389
[10]   Molecular mechanisms of blister formation in bullous impetigo and staphylococcal scalded skin syndrome [J].
Hanakawa, Y ;
Schechter, NM ;
Lin, CY ;
Garza, L ;
Li, H ;
Yamaguchi, T ;
Fudaba, Y ;
Nishifuji, K ;
Sugai, M ;
Amagai, M ;
Stanley, JR .
JOURNAL OF CLINICAL INVESTIGATION, 2002, 110 (01) :53-60