Creatine kinase rabbit muscle

In our laboratory, we have focused our attention on the syntheses of various analogs of creatine, 17, a substrate for the enzyme creatine kinase from rabbit muscle (2). The reaction catalyzed by this enzyme is... 

Wang S, Wang X, Shi W et al (2008) Detection of local polarity and conformational changes at the active site of rabbit muscle creatine kinase with a new arginine-specific fluorescent probe. Biochim Biophys Acta 1784 415 -22... 

In the course of studying the mechanism of action of creatine kinase from rabbit skeletal muscle (M.M isoenzyme), Kenyon and coworkers (4,90) have been involved in the design of specific irreversible inhibitors that are active-site-directed (affinity labels). Creatine kinase catalyzes the reversible transfer of a phosphoryl group ( the elements of "POi") from ATP to creatine, as shown in the following reaction ... 

Hershenson, S. Helmers, N. Desmueles, P. Stroud, R. Purification and crystallization of creatine kinase from rabbit skeletal muscle. J. Biol. Chem., 261, 3732-3736 (1986)... 

Reddy, S.R.R. Watts, D.C. Inhibition of rabbit muscle creatine kinase by iodomethane [proceedings]. Biochem. Soc. Trans., 6, 553-555 (1978)... 

Couthon, R Clottes, E. Vial, C. High salt concentrations induce dissociation of dimeric rabbit muscle creatine kinase. Physico-chemical characterization of the monomeric species. Biochim. Biophys. Acta, 1339, 277-288 (1997)... 

Rao, J.K. Bujacz, G. Wlodawer, A. Crystal structure of rabbit muscle creatine kinase. FEBS Lett., 439, 133-137 (1998)... 

Creatine kinase sequences are known for many different species and iso-forms, so species-specificity of MAbs can often be used for refining the details of epitope mapping. Natural variants that prevent MAb binding are likely to involve contact residues, because the overall protein structures (and enzyme activity) are likely to be retained. The CK-2A7 MAb in Fig. IB binds between Met-29 and Cys-73. It recognizes rabbit and Torpedo CKs, as well as chick CK, but it fails to bind to either rat muscle CK or rabbit brain CK. This suggests that Lys-39 is required for CK-2A7 binding, since it is replaced by Asn m rat muscle CK and by Ala in rabbit brain CK (7), and is the only amino acid change consistent with the observed CK-2A7 specificity. 

Creatine kinase was purified from rabbit muscle by the method of Kuby et al, (4). Rabbit muscle pyruvate kinase was purchased from Boehringer. Porcine muscle adenylate kinase was purchased from Sigma, and was further purified by gel filtration on Sephadex G-50. The enzymes were homogeneous as judged by their specific activities and by their migration as single components in sodium dodecyl sulfate gel electrophoresis. Proton NMR spectra at 250 MHz of 0.5-2.0 mM enzyme sites in 0 solution were obtained with a Bruker WM 250 MHz pulse FT spectrometer at 25°. At least 256 transients were accumulated over 8192 data points using 16 bit A/D conversion. Relaxation rates and histidine pK values were determined by standard NMR methods (5, 6),... 

FIGURE 6.18 In (a) is the MO, and in (b) the 0kl zones of reciprocal space for an orthorhombic crystal of rabbit muscle creatine kinase (space group P212121, a = 47 A, b = 86 A, c = 125 A). Both images exhibit mm symmetry and systematic absences characteristic of 2, axes along both the vertical and horizontal axes. The rotation angle of the crystal between (a) and (b) is 90° about the horizontal axis. 

Liang, Y. Du, F. Sanglier, S. Zhou, B. R. Xia, Y Van Dorsselaer, A. Maechling, C. Kilhoeeer, M. C. Haiech, j. Unfolding of rabbit muscle creatine kinase induced by acid. A study using electrospray ionization mass spectrometry, isothermal titration calorimetry, and fluorescence spectroscopy. J. Biol. Chem. 2003, 27S(32), 30098-30105. 

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