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 language | English |
|---|---|
| Pages (from-to) | 514-519 |
| Number of pages | 6 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 223 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Jun 25 1996 |
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ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
Purification and characterization of recombinant human 5'-methylthioadenosine phosphorylase : Definite identification of coding cDNA. / Della Ragione, Fulvio; Takabayashi, Kenji; Mastropietro, Silvia; Mercurio, Ciro; Oliva, Adriana; Russo, Gian Luigi; Della Pietra, Valentina; Borriello, Adriana; Nobori, Tsumotu; Carson, Dennis A.; Zappia, Vincenzo.
In: Biochemical and Biophysical Research Communications, Vol. 223, No. 3, 25.06.1996, p. 514-519.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification and characterization of recombinant human 5'-methylthioadenosine phosphorylase
T2 - Definite identification of coding cDNA
AU - Della Ragione, Fulvio
AU - Takabayashi, Kenji
AU - Mastropietro, Silvia
AU - Mercurio, Ciro
AU - Oliva, Adriana
AU - Russo, Gian Luigi
AU - Della Pietra, Valentina
AU - Borriello, Adriana
AU - Nobori, Tsumotu
AU - Carson, Dennis A.
AU - Zappia, Vincenzo
PY - 1996/6/25
Y1 - 1996/6/25
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0030601113&partnerID=8YFLogxK
U2 - 10.1006/bbrc.1996.0926
DO - 10.1006/bbrc.1996.0926
M3 - Article
C2 - 8687427
AN - SCOPUS:0030601113
VL - 223
SP - 514
EP - 519
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 3
ER -