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(E.C.2.7.4.14) NOVOCIB's CMK enzyme is a highly pure and active Human recombinant UMP-CMP Kinase which can be used: • to characterize a monophosphorylated nucleotide analogue as a potential substrate of CMK • to quantify UMP, CMP, dUMP or dCMP in a sample • to synthesize enzymatically the di-phosphate form of nucleotide analogue from its monophosphate form (e.g. dFdC-DP) CMK enzyme is available in a lyophilized form. Additional components for CMK assay can also be provided for the measurement of CMK activity through a coupled LDH/PK enzymatic system (reading at 340nm). * Pricing updated September 23rd, 2011. Prices are subject to change without notice. Shipping charges are not included. ** One unit of UMP-CMP kinase converts 1.0 µmole of UMP and ATP to UDP and ADP per minute at pH 7.6 at 25°C, using a coupled enzyme system with PK/LDH. Specific Activity: ≥ 2.5 U/mg protein. For other size, please contact us Click to order or to get further information |
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Synonyms: cytidylate kinase, deoxycytidylate kinase, deoxycytidine monophosphokinase, dCMP kinase, cytidine monophosphate kinase, CMP kinase (CMK, CMPK), uridine monophosphate kinase (UMK, UMPK), uridine monophosphate/cytidine monophosphate kinase, UMP/CMP kinase (UMP/CMPK), CTP:CMP phosphotranferase, ATP:UMP-CMP phosphotransferase, pyrimidine nucleoside monophosphate kinase (YMPK)
NOVOCIB's Human UMP-CMP kinase (CMK) is a recombinant protein of ca. 27kDa (full length 228-aa form(1)) cloned by RT-PCR amplification of mRNA extracted from Huh7 cells (human hepatoma) and expressed in E.coli.
UMP-CMP kinase plays a critical role in supplying cells with nucleotides by catalysing the phosphorylation of CMP, UMP and dCMP to their respective diphosphates. CMK plays also an important role in the activation of cytidine analogues, aracytidine and gemcitabine, a mainstay of leukaemia and lymphoma therapy(2). CMK has a remarkable ability of to phosphorylate L-nucleotides from their monophosphate to diphosphate forms(3) as shown for β-L-2’3’-dideoxy-3’thiacytidine (L-SSdC, 3-TC or lamividune), an anti-HIV and anti-hepatitis B drug.
Crystal structure of open form of human UMP-CMP kinase has been solved recently(4). These data, together with the homology model of enzyme in closed state, provide a structural basis for understanding the substrate specificity of the enzyme and helps to design new nucleoside analogues of higher phosphorylation efficiency.
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UMP-CMP kinase plays a critical role in supplying cells with nucleotides by catalysing the phosphorylation of CMP, UMP and dCMP to their respective diphosphates. CMK plays also an important role in the activation of cytidine analogues, aracytidine and gemcitabine, a mainstay of leukaemia and lymphoma therapy(2). CMK has a remarkable ability of to phosphorylate L-nucleotides from their monophosphate to diphosphate forms(3) as shown for β-L-2’3’-dideoxy-3’thiacytidine (L-SSdC, 3-TC or lamividune), an anti-HIV and anti-hepatitis B drug.
Crystal structure of open form of human UMP-CMP kinase has been solved recently(4). These data, together with the homology model of enzyme in closed state, provide a structural basis for understanding the substrate specificity of the enzyme and helps to design new nucleoside analogues of higher phosphorylation efficiency.
 Unit Definition: One unit of UMP-CMP kinase converts 1.0 µmole of UMP and ATP to UDP and ADP per minute at pH 7.6 at 25°C, using a coupled enzyme system with PK/LDH. Specific Activity: ≥ 2.5 unit/mg protein. Purity: controlled by SDS-PAGE. 
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Assay condition: Enzymatic activity of UMP-CMP kinase is measured by continuous spectrophotometric assays in a coupled lactate dehydrogenase/pyruvate kinase system. Assays are carried out at 37°C, at 50mM Tris-HCl pH7,6; 50mM KCl, 10mM MgCl2, 5mM ATP, 0,1mM NADH, 1mM phosphoenolpyruvate, 1mM DTT, PK 10U/ml, LDH 15U/ml, 380nM CMK. Reaction is followed in an iEMS Reader MF (Labsystems, Finland) microtiter plate reader at 340nm
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