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KMID : 0545120080180020295
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Journal of Microbiology and Biotechnology
2008 Volume.18 No. 2 p.295 ~ p.298
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Overexpression, Purification, and Preliminary X-ray Crystallographic Analysis of Human Brain-Type Creatine Kinase
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Bong Seung-Min
Moon Jin-Ho
Jang Eun-Hyuk
Lee Ki-Seog
Chi Young-Min
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Abstract
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Creatine kinase (CK; E.C. 2.7.3.2) is an important enzyme that catalyzes the reversible transfer of a phosphoryl group from ATP to creatine in energy homeostasis. The brain-type cytosolic isoform of creatine kinase (BB-CK), which is found mainly in the brain and retina, is a key enzyme in brain energy metabolism, because high-energy phosphates are transfered through the creatine kinase/phosphocreatine shuttle system. The recombinant human BB-CK protein was overexpressed as a soluble form in Escherichia coli and crystallized at 22oC using PEG 4000 as a precipitant. Native X-ray diffraction data were collected to 2.2 A resolution using synchrotron radiation. The crystals belonged to the tetragonal space group P43212, with cell parameters of a=b=97.963, c= 164.312 A, and α=β=γ=90o. The asymmetric unit contained two molecules of CK, giving a crystal volume per protein mass (Vm) of 1.80 A3 Da-1 and a solvent content of 31.6%.
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KEYWORD
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Brain-type creatine kinase, shuttle system, energy homeostasis
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