Kagawa T F, Cooney J C, Baker H M, McSweeney S, Liu M, Gubba S, Musser J M, Baker E N
School of Biological Sciences, Department of Chemistry, University of Auckland, Private Bag 92-019, Auckland, New Zealand.
Proc Natl Acad Sci U S A. 2000 Feb 29;97(5):2235-40. doi: 10.1073/pnas.040549997.
Pathogenic bacteria secrete protein toxins that weaken or disable their host, and thereby act as virulence factors. We have determined the crystal structure of streptococcal pyrogenic exotoxin B (SpeB), a cysteine protease that is a major virulence factor of the human pathogen Streptococcus pyogenes and participates in invasive disease episodes, including necrotizing fasciitis. The structure, determined for the 40-kDa precursor form of SpeB at 1.6-A resolution, reveals that the protein is a distant homologue of the papain superfamily that includes the mammalian cathepsins B, K, L, and S. Despite negligible sequence identity, the protease portion has the canonical papain fold, albeit with major loop insertions and deletions. The catalytic site differs from most other cysteine proteases in that it lacks the Asn residue of the Cys-His-Asn triad. The prosegment has a unique fold and inactivation mechanism that involves displacement of the catalytically essential His residue by a loop inserted into the active site. The structure also reveals the surface location of an integrin-binding Arg-Gly-Asp (RGD) motif that is a feature unique to SpeB among cysteine proteases and is linked to the pathogenesis of the most invasive strains of S. pyogenes.
致病细菌分泌的蛋白质毒素会削弱或损害其宿主,从而起到毒力因子的作用。我们已经确定了化脓性链球菌致热外毒素B(SpeB)的晶体结构,它是一种半胱氨酸蛋白酶,是人类病原体化脓性链球菌的主要毒力因子,并参与包括坏死性筋膜炎在内的侵袭性疾病发作。以1.6埃分辨率确定的40 kDa前体形式的SpeB结构表明,该蛋白质是木瓜蛋白酶超家族的远亲同源物,该超家族包括哺乳动物组织蛋白酶B、K、L和S。尽管序列同一性可忽略不计,但蛋白酶部分具有典型的木瓜蛋白酶折叠结构,尽管有主要的环插入和缺失。催化位点与大多数其他半胱氨酸蛋白酶不同,因为它缺乏Cys-His-Asn三联体中的Asn残基。前肽具有独特的折叠和失活机制,该机制涉及通过插入活性位点的环取代催化必需的His残基。该结构还揭示了整合素结合Arg-Gly-Asp(RGD)基序的表面位置,这是SpeB在半胱氨酸蛋白酶中独有的特征,并且与化脓性链球菌最具侵袭性菌株的发病机制有关。