Protein Expr Purif. 2000 Dec;20(3):365-71.

Rational design and purification of human Bruton's tyrosine kinase SH3-SH2 protein for structure-function studies.

Nera KP, Brockmann E, Vihinen M, Smith CI, Mattsson PT.

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Department of Biochemistry and Food Chemistry, University of Turku, FIN-20014 Turku, Finland.

Abstract

Bruton's tyrosine kinase (Btk) is a cytoplasmic protein tyrosine kinase consisting of N-terminal pleckstrin homology (PH) domain followed by Tec homology (TH) domain, Src homology 3 and 2 (SH3 and SH2) domains, and a C-terminal kinase domain. Mutations in the human BTK gene cause the severe immunodeficiency disease X-linked agammaglobulinemia (XLA). The structural and functional basis of several XLA-causing mutations remains unknown, since only the structures of the PH and SH3 domains of human Btk are currently available. In this study, we overexpressed and purified a protein consisting of the SH3 and SH2 domains of human Btk for biochemical and structural analysis. The purified protein was only partially soluble and had a tendency to dimerize, which made it unsuitable for further studies. To overcome the problems of low solubility and dimerization, subdomain interactions were engineered without altering the function of the protein.

PMID:: 11087675; [PubMed - indexed for MEDLINE]

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Six X-linked agammaglobulinemia-causing missense mutations in the Src homology 2 domain of Bruton's tyrosine kinase: phosphotyrosine-binding and circular dichroism analysis. [J Immunol. 2000]
Six X-linked agammaglobulinemia-causing missense mutations in the Src homology 2 domain of Bruton's tyrosine kinase: phosphotyrosine-binding and circular dichroism analysis.
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Deletion within the Src homology domain 3 of Bruton's tyrosine kinase resulting in X-linked agammaglobulinemia (XLA).
Zhu Q, Zhang M, Rawlings DJ, Vihinen M, Hagemann T, Saffran DC, Kwan SP, Nilsson L, Smith CI, Witte ON, et al. J Exp Med. 1994 Aug 1; 180(2):461-70.
Review Bruton tyrosine kinase (BTK) in X-linked agammaglobulinemia (XLA). [Front Biosci. 2000]
Review Bruton tyrosine kinase (BTK) in X-linked agammaglobulinemia (XLA).
Vihinen M, Mattsson PT, Smith CI. Front Biosci. 2000 Dec 1; 5:D917-28. Epub 2000 Dec 1.
Review BTK, the tyrosine kinase affected in X-linked agammaglobulinemia. [Front Biosci. 1997]
Review BTK, the tyrosine kinase affected in X-linked agammaglobulinemia.
Vihinen M, Mattsson PT, Smith CI. Front Biosci. 1997 Jan 1; 2:d27-42. Epub 1997 Jan 1.

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Cited by 1 PubMed Central article

Conformation of full-length Bruton tyrosine kinase (Btk) from synchrotron X-ray solution scattering. [EMBO J. 2003]
Conformation of full-length Bruton tyrosine kinase (Btk) from synchrotron X-ray solution scattering.
Márquez JA, Smith CI, Petoukhov MV, Lo Surdo P, Mattsson PT, Knekt M, Westlund A, Scheffzek K, Saraste M, Svergun DI. EMBO J. 2003 Sep 15; 22(18):4616-24.

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