Document Detail

Purification and characterization of the nuclear cytidine 5'-monophosphate N-acetylneuraminic acid synthetase from rat liver.
MedLine Citation:
PMID:  1577759     Owner:  NLM     Status:  MEDLINE    
N-Acetylneuraminic acid cytidylyltransferase (EC (CAMP-NeuAc synthetase) from rat liver catalyzes the formation of cytidine monophosphate N-acetylneuraminic acid from CTP and NeuAc. We have purified this enzyme to apparent homogeneity (241-fold) using gel filtration on Sephacryl S-200 and two types of affinity chromatographies (Reactive Brown-10 Agarose and Blue Sepharose CL-6B columns). The pure enzyme, whose amino acid composition and NH2-terminal amino acid sequence are also established, migrates as a single protein band on non-denaturing polyacrylamide gel electrophoresis. The molecular mass of the native enzyme, estimated by gel filtration, was 116 +/- 2 kDa whereas its Mr in sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 58 +/- 1 kDa. CMP-NeuAc synthetase requires Mg2+ for catalysis although this ion can be replaced by Mn2+, Ca2+, or Co2+. The optimal pH was 8.0 in the presence of 10 mM Mg2+ and 5 mM dithiothreitol. The apparent Km for CTP and NeuAc are 1.5 and 1.3 mM, respectively. The enzyme also converts N-glycolylneuraminic acid to its corresponding CMP-sialic acid (Km, 2.6 mM), whereas CMP-NeuAc, high CTP concentrations, and other nucleotides (CDP, CMP, ATP, UTP, GTP, and TTP) inhibited the enzyme to different extents.
L B Rodríguez-Aparicio; J M Luengo; C González-Clemente; A Reglero
Related Documents :
8595079 - Pharmacodynamics of a liposomal preparation for local anaesthesia.
735639 - Purification of acid proteinases from calf brain.
12711419 - Isolation and characterisation of metallothionein from the clam ruditapes decussatus.
23180549 - Heterologous expression and biochemical characterization of acetyl xylan esterase from ...
17157499 - Qualitative analysis of products formed during the acid catalyzed liquefaction of bagas...
6313719 - New ion exchanger for the separation of proteins and nucleic acids.
7150619 - Cyanide-insensitive and clofibrate enhanced beta-oxidation of dodecanedioic acid in rat...
19629599 - Variations in allelochemical composition of leachates of different organs and maturity ...
7055559 - Transport of neutral amino acids by human erythrocytes.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  267     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1992 May 
Date Detail:
Created Date:  1992-06-05     Completed Date:  1992-06-05     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  9257-63     Citation Subset:  IM    
Departamento de Bioquímica y Biología Molecular, Universidad de León, Spain.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Amino Acid Sequence
Amino Acids / analysis
Cell Nucleus / enzymology*
Chromatography, Gel
Enzyme Stability
Liver / enzymology*,  ultrastructure
Molecular Sequence Data
N-Acylneuraminate Cytidylyltransferase / isolation & purification,  metabolism*
Rats, Inbred Strains
Substrate Specificity
Reg. No./Substance:
0/Amino Acids; EC Cytidylyltransferase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  A synthetic peptide derived from p34cdc2 is a specific and efficient substrate of src-family tyrosin...
Next Document:  Sequence requirements for maturation of the 5' terminus of human 18 S rRNA in vitro.