Microsequencing, analysis of molecular weight and amino acid composition for pyrimidine 5'-nucleotidase I of human erythrocytes.
- Author:
Zhu-Lin PAN
1
;
Jin-Ying LI
;
Bi-He MIN
;
Kang YING
;
Hong ZHOU
;
Xiao-Ping XU
;
Xian-Min SHONG
;
Feng-Lai HAN
;
Wei-Ping ZHANG
;
Xian ZHANG
Author Information
1. Department of Hematology, Changhai Hospital, The Second Military Medical University, China. paradise806@sohu.com
- Publication Type:Journal Article
- MeSH:
5'-Nucleotidase;
blood;
chemistry;
isolation & purification;
Amino Acid Sequence;
Amino Acids;
analysis;
Chromatography, High Pressure Liquid;
Electrophoresis, Polyacrylamide Gel;
Erythrocytes;
enzymology;
Humans;
Mass Spectrometry;
Molecular Weight;
Peptide Fragments;
chemistry;
Sequence Analysis, Protein
- From:
Journal of Experimental Hematology
2003;11(1):61-65
- CountryChina
- Language:Chinese
-
Abstract:
To further explore the mechanism of congenital pyrimidine 5'-nuleotidase I (P5'N-I) deficiency, on the basis of purification of the protein, the molecular weight and amino acid composition were analysed by mass-spectrograph and amino-acid analyzer, microsequencing and bioinformation analysis of P5'N-I were performed after it was hydrolysed by trypsin. The results showed that three fractions were found in the purified P5'N-I and their molecular weights were 26,952.9, 55,476 and 110,938, respectively. The sequence from one of the peptide fragments was I-E-G-P-T-I-R-Q-I-E. The homologous sequence was not found after comparision with the ten-amino-acid sequence in GenBank by blast procedure. Amino acid analysis indicated that P5'N-I was composed of 18 amino acids at least, and 243 amino acid residues. In conclusion, the enzyme might be an allosteric enzyme, there might be homologous dimer or tetramer in physiological status of normal human erythrocyte, the microsequence could be designed as the probe for fishing the genes of interest. The composition of amino acid might be an important information in determination of its protein primary structure.
