Purification and characterization of recombinant human 5'-methylthioadenosine phosphorylase: Definite identification of coding cDNA

Fulvio Della Ragione, Kenji Takabayashi, Silvia Mastropietro, Ciro Mercurio, Adriana Oliva, Gian Luigi Russo, Valentina Della Pietra, Adriana Borriello, Tsumotu Nobori, Dennis A. Carson, Vincenzo Zappia

Research output: Contribution to journalArticle

Abstract

5'-Methylthioadenosine phosphorylase gene maps on the 9p21 chromosome, strictly linked to the important tumor suppressor gene p16(INK4A). Chromosomal deletions encompassing both the phosphorylase and p16(INK4A) 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.

Original languageEnglish
Pages (from-to)514-519
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume223
Issue number3
DOIs
Publication statusPublished - Jun 25 1996

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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    Della Ragione, F., Takabayashi, K., Mastropietro, S., Mercurio, C., Oliva, A., Russo, G. L., Della Pietra, V., Borriello, A., Nobori, T., Carson, D. A., & Zappia, V. (1996). Purification and characterization of recombinant human 5'-methylthioadenosine phosphorylase: Definite identification of coding cDNA. Biochemical and Biophysical Research Communications, 223(3), 514-519. https://doi.org/10.1006/bbrc.1996.0926