Six X-linked agammaglobulinemia-causing missense mutations in the src homology 2 domain of Bruton's tyrosine kinase:: Phosphotyrosine-binding and circular dichroism analysis

被引:27
作者
Mattsson, PT [1 ]
Lappalainen, I
Bäckesjö, CM
Brockmann, E
Laurén, S
Vihinen, M
Smith, CIE
机构
[1] Univ Turku, Dept Biochem & Food Chem, FIN-20014 Turku, Finland
[2] Huddinge Univ Hosp, Karolinska Inst, Ctr Biotechnol, Dept Biosci, S-14186 Huddinge, Sweden
[3] Huddinge Univ Hosp, Karolinska Inst, Dept Immunol Microbiol Pathol & Infect Dis, IMPI, S-14186 Huddinge, Sweden
[4] Univ Helsinki, Dept Biosci, Div Biochem, Helsinki, Finland
[5] Univ Tampere, Inst Med Technol, FIN-33101 Tampere, Finland
关键词
D O I
10.4049/jimmunol.164.8.4170
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Src homology 2 (SH2) domains recognize phosphotyrosine (pY)-containing sequences and thereby mediate their association to ligands. Bruton's tyrosine kinase (Btk) is a cytoplasmic protein tyrosine kinase, in which mutations cause a hereditary immunodeficiency disease, X-linked agammaglobulinemia (XLA). Mutations have been found in all Btk domains, including SH2, We have analyzed the structural and functional effects of six disease-related amino acid substitutions in the SH2 domain: G302E, R307G, Y334S, L358F, Y361C, and H362Q, Also, we present a novel Btk SH2 missense mutation, H362R, leading to classical XLA, Based on circular dichroism analysis, the conformation of five of the XLA mutants studied differs from the native Btk SH2 domain, while mutant R307G is structurally identical, The binding of XLA mutation-containing SH2 domains to pY-Sepharose was reduced, varying between 1 and 13% of that for the native SH2 domain. The solubility of all the mutated proteins was remarkably reduced. SH2 domain mutations were divided into three categories: 1) Functional mutations, which affect residues presumably participating directly in pY binding (R307G); 2) structural mutations that, via conformational change, not only impair pY binding, but severely derange the structure of the SH2 domain and possibly interfere with the overall conformation of the Btk molecule (G302E, Y334S, L358F, and H362Q); and 3) structural-functional mutations, which contain features from both categories above (Y361C).
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页码:4170 / 4177
页数:8
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