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Updated: Aug 13, 2026
Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions
Published on: February 10, 2022
Stoichiometry of lipid-protein interaction assessed by hydrophobic photolabeling
Ana María Villamil Giraldo1, Pablo Raúl Castello, F Luis González Flecha
1Instituto de Química y Fisicoquímica Biológicas, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956.C1113AAD, Buenos Aires, Argentina.
Abstract:
Here we undertook a comparative study of the composition of the lipid annulus of three ATPases pertaining to the P-type family: plasma membrane calcium pump (PMCA), sarcoplasmic reticulum calcium pump (SERCA) and Na,K-ATPase. The photoactivatable phosphatidylcholine analogue [(125)I]TID-PC/16 was incorporated into mixtures of dimyristoyl phosphatidylcholine (DMPC) and each enzyme with the aid of the nonionic detergent C(12)E(10). After photolysis, the extent of the labeling reaction was assessed to determine the lipid:protein stoichiometry: 17 for PMCA, 18 for SERCA, 24 for the Na,K-ATPase (alpha-subunit) and 5.6 mol PC/mol protein for the Na,K-ATPase (beta-subunit).
