The muscle fatty acid binding protein of spadefoot toad (Scaphiopus couchii).
Fatty acid binding protein was purified from skeletal muscle of the spadefoot toad (Scaphiopus couchii), an estivating species. While estivating, this animal relies on the fatty acid oxidation for energy. Hence we were interested in the behaviour of fatty acid binding protein under conditions of elevated urea (up to 200 mM) and potassium chloride such as exist during estivation. Also we examined whether there were interactions between glycolytic intermediates and the binding ability of the protein. The amount of bound fatty acid (a fluorescence assay using cis-parinarate) was not affected (P < 0.05) by glucose, fructose 6-phosphate or phosphoenolpyruvate at physiological concentrations. By contrast, glucose 6-phosphate increased the amount of bound cis-parinarate but the apparent dissociation constant was not different from the control. Fructose 1,6-bisphosphate but not fructose 2,6-phosphate decreased cis-parinarate binding by 40%, commensurate with doubling the apparent dissociation constant (1.15-2.62 microM). Urea, guanidinium and trimethylamine N-oxide at 200 mM increased cis-parinarate binding 60% over controls. Urea (1 M) and KCl (200 mM) did not affect cis-parinarate binding compared to controls. The interaction of this fatty acid transporter with fructose 1,6-bisphosphate is discussed in terms of reciprocal interaction with phosphofructokinase since fatty acid is also an inhibitor of phosphofructokinase.
Stewart
J M
JM
Department of Biology, Mount Allison University, Sackville, NB, Canada. jstewart@mta.ca
Claude
J F
JF
MacDonald
J A
JA
Storey
K B
KB
D000818:Animals; D001001:Anura / Q000502:physiology*; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004956:Estivation / Q000502:physiology; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids / Q000378:metabolism*; D005456:Fluorescent Dyes; D005947:Glucose / Q000494:pharmacology; D006019:Glycolysis; D006600:Hexosephosphates / Q000494:pharmacology; D007700:Kinetics; D009124:Muscle Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D009363:Neoplasm Proteins; D019301:Oleic Acid / Q000494:pharmacology; D009994:Osmolar Concentration; D010728:Phosphoenolpyruvate / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D014508:Urea / Q000378:metabolism
D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D005456:Fluorescent Dyes; D006600:Hexosephosphates; D009124:Muscle Proteins; D009363:Neoplasm Proteins; D019301:Oleic Acid; D010728:Phosphoenolpyruvate; D014508:Urea; D005947:Glucose