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PMID:8687427

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Citation

Della Ragione, F, Takabayashi, K, Mastropietro, S, Mercurio, C, Oliva, A, Russo, GL, Della Pietra, V, Borriello, A, Nobori, T, Carson, DA and Zappia, V (1996) Purification and characterization of recombinant human 5'-methylthioadenosine phosphorylase: definite identification of coding cDNA. Biochem. Biophys. Res. Commun. 223:514-9

Abstract

5'-Methylthioadenosine phosphorylase gene maps on the 9p21 chromosome, strictly linked to the important tumor suppressor gene p16INK4A. Chromosomal deletions encompassing both the phosphorylase and p16INK4A genes cause the complete absence of the enzymatic activity in a large number of tumors, thus resulting in well-defined metabolic differences between malignant and normal cells. Recently, the cloning of the phosphorylase gene has been reported on the basis of indirect evidence. In order to demonstrate definitely the identification of 5'-methylthioadenosine phosphorylase gene, we have cloned the putative enzyme coding sequence in a prokaryotic expression vector and expressed the protein in bacteria. The recombinant phosphorylase has been purified to homogeneity and its physicochemical, immunological and kinetic features have been characterized. The results obtained allowed the conclusive demonstration of 5'-methylthioadenosine phosphorylase gene cloning and the use of recombinant protein for further characterization.

Links

PubMed Online version:10.1006/bbrc.1996.0926

Keywords

Chromatography, Gel; Chromatography, Ion Exchange; Chromosome Mapping; Chromosomes, Human, Pair 9; Cloning, Molecular; DNA, Complementary; Enzyme Inhibitors/pharmacology; Escherichia coli; Genetic Linkage; Humans; Isopropyl Thiogalactoside/pharmacology; Molecular Weight; Phosphates/pharmacology; Purine-Nucleoside Phosphorylase/genetics; Purine-Nucleoside Phosphorylase/isolation & purification; Purine-Nucleoside Phosphorylase/metabolism; Recombinant Proteins/biosynthesis; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism; Substrate Specificity

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:MTAP

enables

GO:0004645: 1,4-alpha-oligoglucan phosphorylase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:MTAP

enables

GO:0004645: phosphorylase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:MTAP

involved_in

GO:0006139: nucleobase-containing compound metabolic process

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete


See also

References

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