Mercury-induced crystallization and SAD phasing of the human Fe65-PTB1 domain.
Fe65 is a three-domain neuronal adaptor protein involved in brain development and amyloid precursor protein (APP) signalling. The phosphotyrosine-binding domain 1 (PTB1) of human Fe65 has been cloned, overexpressed, purified and crystallized using the hanging-drop vapour-diffusion method. Native crystals belong to the space group R3 and diffract to 2.6 A resolution. This crystal form suffered from high thermal B factors and pseudo-symmetry, resulting in a bisection of the c axis. Co-crystallization with a mercury compound under similar conditions induced an orthorhombic crystal form in the space group P2(1)2(1)2(1) diffracting to 2.2 A resolution. SAD phases have been computed to the diffraction limit at the wavelength of maximum absorption (L(III) edge).
Radzimanowski
Jens
J
Heidelberg University Biochemistry Center, INF328, D-69120 Heidelberg, Germany.
Ravaud
Stéphanie
S
Beyreuther
Konrad
K
Sinning
Irmgard
I
Wild
Klemens
K
D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D008628:Mercury / Q000494:pharmacology*; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D019000:Phosphotyrosine / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary
C091042:APBB1 protein, human; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D019000:Phosphotyrosine; D008628:Mercury