Use this URL to cite or link to this record in EThOS:
Title: Studies on the structure of human platelet phosphofructokinase
Author: Simpson, Craig J.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1991
Availability of Full Text:
Access from EThOS:
Full text unavailable from EThOS. Please try the link below.
Access from Institution:
Phosphofructokinase (ATP: D-fructose 6-phosphate 1-transferase, EC catalyses the phosphorylation of fructose 6-phosphate to fructose 1,6-bisphosphate, and is a key regulatory enzyme in glycolysis. Sequence conservation between mammalian PFK and PFK from E.coli, B.stearothermophilus and S.citri, and between the amino and carboxyl halves of the mammalian enzymes suggests that mammalian PFK has evolved from a prokaryotic progenitor by a process of gene duplication and fusion. The aim of this project was to obtain detailed structural information describing PFK from humans and from the parasitic filarial nematode Onchocerca volvulus and so facilitate the design of inhibitors specific for the filarial enzyme. Five Onchocerca volvulus cDNA libraries were probed with oligonucleotides, heterologous DNA from E.coli, rabbit muscle and human muscle PFK. Redundant oligonucleotides were used in a polymerase chain reaction to try to clone part of the parasite's PFK. Antisera to E.coli and Ascaris suum PFK were used to screen libraries for expression of PFK. The lack of good quality DNA libraries and the unexpected gene structure and codon usage of the parasite did not allow the isolation of DNA encoding O.volvulus PFK. In man, PFK is under the control of three structural loci encoding subunits for muscle, liver and platelet types. They are located on separate chromosomes and are regulated independently. During the course of this project, the muscle and liver type DNA sequences were reported from other laboratories. The partial sequence of the platelet type is reported here, and its chromosomal location is confirmed. The sequence codes for three hundred amino acids and comprises 80% of the carboxyl domain in comparison to the muscle and liver types. The structure, function and evolution of PFK is discussed, based on detailed crystal structure information of PFK from E.coli and B.stearothermophilus and the amino acid sequences of PFK from other sources.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available