Expression of pyrimidine 5'-nucleotidase subclass I during erythrocyte maturation in rats.
The subclass I enzyme of rat pyrimidine 5'-nucleotidase (P5N-I), which preferentially hydrolyzes (deoxy)CMP and UMP, is distributed specifically in red blood cells (RBCs), and its activity increases approximately six-fold as compared to the control value after erythropoietic induction by phenylhydrazine administration. In this study, we detected rat P5N-I protein by using antibodies against the chicken P5N-I enzyme. The molecular mass of rat P5N-I was approximately 37 kDa, as estimated by gel filtration chromatography and Western blot analysis. The pI value of the enzyme was approximately 5.7. This protein band was detected only in RBC lysate extract, i.e., not in cytosol from the erythropoietic spleen. Protein mass of the P5N-I enzyme, estimated by immunoblot analysis, was increased in proportion to the enzyme activity after erythropoietic induction in rats. No phosphorylation of the enzyme protein was detected by immunoblot analysis with anti-phosphoserine or anti-phosphotyrosine antibody. In conclusion, these findings indicate that the rat P5N-I enzyme is expressed specifically in reticulocytes and may therefore be essential in the maturation process of rat erythrocytes.
Hokari
S
S
Department of Biochemistry, Junior College, Saitama Medical School, Iruma-gun, Saitama 350- 0495, Japan.
Miyazaki
T
T
Matsunaga
T
T
Koyama
I
I
Hasegawa
M
M
Komoda
T
T
D015720:5'-Nucleotidase / Q000096:biosynthesis* / Q000302:isolation & purification; D000818:Animals; D015153:Blotting, Western; D004912:Erythrocytes / Q000201:enzymology / Q000502:physiology*; D004920:Erythropoiesis / Q000187:drug effects; D010659:Phenylhydrazines / Q000494:pharmacology; D051381:Rats; D012156:Reticulocytes / Q000201:enzymology; D013997:Time Factors
D010659:Phenylhydrazines; C030299:phenylhydrazine; D015720:5'-Nucleotidase