Overexpression, purification, and preliminary X-ray crystallographic analysis of human brain-type creatine kinase

Seung Min Bong, Jin H. Moon, Eun Hyuk Jang, Ki Seog Lee, Young Min Chi

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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 22°C using PEG 4000 as a precipitant. Native X-ray diffraction data were collected to 2.2 Å resolution using synchrotron radiation. The crystals belonged to the tetragonal space group P43212, with cell parameters of a=b=97.963, c=164.312 Å, and α= β=γ=90°. The asymmetric unit contained two molecules of CK, giving a crystal volume per protein mass (Vm) of 1.80 Å3 Da-1 and a solvent content of 31.6%.

Original languageEnglish
Pages (from-to)295-298
Number of pages4
JournalJournal of Microbiology and Biotechnology
Volume18
Issue number2
Publication statusPublished - 2008 Feb 28

Fingerprint

BB Form Creatine Kinase
Purification
Brain
X-Rays
Creatine Kinase
X rays
Enzymes
Proteins
Crystals
Synchrotrons
Phosphocreatine
Creatine
Adenosinetriphosphate
Energy Transfer
Synchrotron radiation
X-Ray Diffraction
Escherichia coli
Energy Metabolism
Polyethylene glycols
Retina

Keywords

  • Brain-type creatine kinase
  • Energy homeostasis
  • Shuttle system

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Overexpression, purification, and preliminary X-ray crystallographic analysis of human brain-type creatine kinase. / Bong, Seung Min; Moon, Jin H.; Jang, Eun Hyuk; Lee, Ki Seog; Chi, Young Min.

In: Journal of Microbiology and Biotechnology, Vol. 18, No. 2, 28.02.2008, p. 295-298.

Research output: Contribution to journalArticle

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