Crystallization and preliminary crystallographic analysis of the motor domain of human kinetochore-associated protein CENP-E using an automated crystallization procedure.
Human centromere-associated protein E, a member of the kinesin superfamily, is a microtubule-dependent motor protein involved in cell division that has been localized transiently to the kinetochore. The protein is thought to be responsible for the correct attachment and positioning of chromosomes to the mitotic spindle during the metaphase. The 312 kDa protein comprises four different domains. In this study, the focus was on the N-terminal motor domain, which includes the ATP-binding site and a region for microtubule binding. Crystals of the CENP-E motor domain have been obtained by high-throughput crystallization screening using an automated TECAN crystallization robot. The crystals (737 x 132 x 79 microm) belong to the space group P2(1), with unit-cell parameters a = 49.35, b = 83.70, c = 94.16 angstroms, beta = 103.05 degrees. They diffract to 2.1 angstroms resolution using synchrotron radiation.
Garcia-Saez
Isabel
I
Laboratoire de Microscopie Electronique Structurale, Institut de Biologie Structurale Jean-Pierre Ebel (CEA-CNRS-UJF), 41 Rue Jules Horowitz, 38027 Grenoble CEDEX 01, France. isabel.garcia@ibs.fr
Blot
Delphine
D
Kahn
Richard
R
Kozielski
Frank
F
D000255:Adenosine Triphosphate / Q000737:chemistry; D016880:Anisotropy; D001665:Binding Sites; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry*; D002875:Chromosomes / Q000648:ultrastructure; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004247:DNA / Q000737:chemistry; D006801:Humans; D008938:Mitosis; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D008941:Spindle Apparatus / Q000378:metabolism
D002868:Chromosomal Proteins, Non-Histone; D011994:Recombinant Proteins; C068688:centromere protein E; D000255:Adenosine Triphosphate; D004247:DNA