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The effect of lipids on the enzymatic activity of 6-phosphofructo-1-kinase from B. stearothermophilus

The effect of lipids on the enzymatic activity of 6-phosphofructo-1-kinase from B. stearothermophilus
The effect of lipids on the enzymatic activity of 6-phosphofructo-1-kinase from B. stearothermophilus
6-Phosphofructo-1-kinase (PFK-1), a major regulatory enzyme in the glycolysis pathway, is a cytoplasmic enzyme with complicated allosteric kinetics. Here we investigate the effects of lipids on the activity of PFK from Bacillus stearothermophilus (BsPFK), to determine whether BsPFK shares any of the membrane binding or lipid binding properties reported for some mammalian PFKs. Our results show that large unilamellar vesicles (LUVs) composed of either the phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or of 1:1 (mole ratio) DOPC and the fatty acid, oleic acid (OA), cause a three-fold increase in Vmax, depending on the lipid concentration and vesicle composition, but no change in Km. Further studies show lipids do not reverse the allosteric inhibitory effects of phosphoenolpyruvate (PEP) on BsPFK. SDS/PAGE studies do not show significant binding of the BsPFK tetramer to the surface of the phospholipid vesicles, suggesting that modulation of catalytic activity is due to binding of lipid monomers. By simulating the kinetics of BsPFK interaction with vesicles and lipid monomers we conclude that the change in BsPFK catalytic activity with respect to lipid concentration is consistent with monomer abstraction from vesicles rather than direct uptake of lipid monomers from solution.
0009-3084
Tsaloglou, Maria-Nefeli
99ab30ba-15da-4d25-86ba-608d127f8369
Attard, George S.
3219075d-2364-4f00-aeb9-1d90f8cd0d36
Dymond, Marcus K.
e180765b-039e-47ff-9841-ce6a5123a519
Tsaloglou, Maria-Nefeli
99ab30ba-15da-4d25-86ba-608d127f8369
Attard, George S.
3219075d-2364-4f00-aeb9-1d90f8cd0d36
Dymond, Marcus K.
e180765b-039e-47ff-9841-ce6a5123a519

Tsaloglou, Maria-Nefeli, Attard, George S. and Dymond, Marcus K. (2011) The effect of lipids on the enzymatic activity of 6-phosphofructo-1-kinase from B. stearothermophilus. Chemistry and Physics of Lipids. (doi:10.1016/j.chemphyslip.2011.08.003).

Record type: Article

Abstract

6-Phosphofructo-1-kinase (PFK-1), a major regulatory enzyme in the glycolysis pathway, is a cytoplasmic enzyme with complicated allosteric kinetics. Here we investigate the effects of lipids on the activity of PFK from Bacillus stearothermophilus (BsPFK), to determine whether BsPFK shares any of the membrane binding or lipid binding properties reported for some mammalian PFKs. Our results show that large unilamellar vesicles (LUVs) composed of either the phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or of 1:1 (mole ratio) DOPC and the fatty acid, oleic acid (OA), cause a three-fold increase in Vmax, depending on the lipid concentration and vesicle composition, but no change in Km. Further studies show lipids do not reverse the allosteric inhibitory effects of phosphoenolpyruvate (PEP) on BsPFK. SDS/PAGE studies do not show significant binding of the BsPFK tetramer to the surface of the phospholipid vesicles, suggesting that modulation of catalytic activity is due to binding of lipid monomers. By simulating the kinetics of BsPFK interaction with vesicles and lipid monomers we conclude that the change in BsPFK catalytic activity with respect to lipid concentration is consistent with monomer abstraction from vesicles rather than direct uptake of lipid monomers from solution.

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More information

Published date: 17 August 2011
Organisations: Chemistry, Ocean Technology and Engineering

Identifiers

Local EPrints ID: 195535
URI: http://eprints.soton.ac.uk/id/eprint/195535
ISSN: 0009-3084
PURE UUID: 1604c00a-a529-4a8e-a9ec-b84669c85536
ORCID for George S. Attard: ORCID iD orcid.org/0000-0001-8304-0742

Catalogue record

Date deposited: 22 Aug 2011 16:13
Last modified: 20 Jul 2019 01:21

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Contributors

Author: Maria-Nefeli Tsaloglou
Author: Marcus K. Dymond

University divisions

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