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. 1974 Sep;141(3):753–760. doi: 10.1042/bj1410753

Studies of triose phosphate isomerase by hydrogen exchange

Christopher A Browne 1, Stephen G Waley 1
PMCID: PMC1168182  PMID: 4478068

Abstract

The 3H–H exchange of chicken muscle and rabbit muscle triose phosphate isomerases was studied. Their behaviour was mostly very similar. `Exchange-in' (acquisition of radioactivity when protein was incubated in 3H2O) was measured at 37°C and at pH7.5, and the rates of exchange of the native and liganded enzymes were compared. Inhibitors and substrates retarded exchange, substrates showing the most marked effect; structural rearrangements in the enzyme may thus play some part in catalysis. The inhibitor phosphoglycollate affected the rabbit enzyme, but had little or no effect on the chicken enzyme. `Exchange-out' (loss of radioactivity from protein previously labelled by incubation in 3H2O) was measured by hollow-fibre dialysis. When ligand was removed during the course of dialysis (by replacing buffer that contained ligand with buffer that lacked ligand) there was a prompt decrease in the number of labelled H atoms of the protein. Analysis of the curves provides some information about the number and half-lives of the responsive H atoms. Ligands decrease the motility of the protein and affect about one-fifth of the chain. Low concentrations of glycerol 3-phosphate have an effect that is greater than expected.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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