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Isolation and sequence of a cDNA encoding human platelet phosphofructokinase
C J Simpson1, L A Fothergill-Gilmore
1Department of Biochemistry, University of Edinburgh, Scotland.
Insights
Researchers isolated human platelet 6-phosphofructokinase (PFK) cDNA, revealing high amino acid similarity to muscle and liver PFK isoenzymes. The platelet PFK gene was mapped to chromosome 10p15.2-p15.3.
Area of Science:
- Biochemistry
- Molecular Biology
- Human Genetics
Background:
- 6-phosphofructokinase (PFK) is a key glycolytic enzyme with multiple isoenzymes.
- Understanding the structure and genetic location of platelet PFK is crucial for metabolic studies.
Purpose of the Study:
- To isolate and characterize the cDNA encoding human platelet 6-phosphofructokinase (PFK).
- To determine the sequence identity and conserved functional sites of platelet PFK compared to other isoenzymes.
- To map the genetic locus of the human platelet PFK gene.
Main Methods:
- cDNA library screening using a human muscle PFK probe.
- DNA sequencing to determine the coding and untranslated regions.
- Amino acid sequence alignment to compare isoenzymes.
- Chromosomal mapping using biotinylated cDNA probes.
Main Results:
- Isolation of a human platelet PFK cDNA clone.
- Deduced amino acid sequence shows 71% identity to muscle and 63% identity to liver PFK.
- Catalytic and effector sites are conserved across isoenzymes.
- The human platelet PFK gene is assigned to chromosome 10p15.2-p15.3.
Conclusions:
- The human platelet PFK isoenzyme shares significant sequence homology with other PFK forms.
- Conserved functional sites suggest similar enzymatic mechanisms.
- The localization of the platelet PFK gene provides a basis for further genetic and functional studies.
Abstract:
A cDNA encoding human platelet 6-phosphofructokinase (PFK; EC 2.7.1.11) has been isolated from a human lymphocyte Raji cell line cDNA library using a cDNA for human muscle PFK as a probe. The platelet cDNA contains 900bp of carboxy terminal coding sequence and 238bp of downstream untranslated region. The deduced amino acid sequence shows 71% identity to the amino acid sequence for the human muscle isoenzyme and 63% identity to the human liver isoenzyme. Almost all of the amino acid residues contributing to catalytic and effector sites in the three isoenzymes are conserved. The platelet gene has been assigned to chromosome 10p15.2-p15.3 by using the cDNA clone as a biotinylated probe against human chromosome spreads (Morrison et al. 1991, submitted to Human Genetics).
