cDNA encoding type B subunit of rat phosphoglycerate mutase: its isolation and nucleotide sequence

Area of Science:

• Biochemistry
• Molecular Biology
• Genetics

Background:

• Phosphoglycerate mutase (PGAM) is a key enzyme in glycolysis.
• The nonmuscle-specific subunit (PGAM-B) plays a crucial role in cellular energy metabolism.

Purpose of the Study:

• To isolate and characterize the cDNA encoding the nonmuscle-specific (type B) subunit of phosphoglycerate mutase (PGAM-B) in rats.
• To compare the deduced amino acid sequence of rat PGAM-B with its human counterpart.

Main Methods:

• Synthesis of a cDNA probe using polymerase chain reaction (PCR) from rat liver cell mRNA.
• Screening of a rat liver cell cDNA library using the synthesized probe.
• Confirmation of cDNA identity via amino acid sequencing of purified protein peptides.
• RNA blot analysis to determine mRNA size.

Main Results:

• Isolation and characterization of a cDNA encoding rat PGAM-B.
• The coding region specifies a polypeptide of 253 amino acids plus initiator methionine.
• RNA blot analysis revealed a single mRNA species of 1.7 kilobases in rat liver cells.
• The deduced amino acid sequence of rat PGAM-B showed high identity to human PGAM-B, with a single substitution (Valine for Rat, Alanine for Human) at position 251.

Conclusions:

• The study successfully isolated and characterized the rat PGAM-B cDNA.
• Rat and human PGAM-B share significant sequence homology, indicating conserved function.
• The findings contribute to understanding the molecular basis of PGAM-B in mammals.