Lack of activation of UCP1 in isolated brown adipose tissue mitochondria by glucose-O-ω-modified saturated fatty acids of various chain lengths

Abstract

We previously demonstrated that uncoupling protein 1 activity, as measured in isolated brown adipose tissue mitochondria (and as a native protein reconstituted into liposome membranes), was not activated by the non-flippable modified saturated fatty acid, glucose-O-ω-palmitate, whereas activity was stimulated by palmitate alone (40 nM free final concentration). In this study, we investigated whether fatty acid chain length had any bearing on the ability of glucose-O-ω-fatty acids to activate uncoupling protein 1. Glucose-O-ω-saturated fatty acids of various chain lengths were synthesized and tested for their potential to activate GDP-inhibited uncoupling protein 1-dependent oxygen consumption in brown adipose tissue mitochondria, and the results were compared with equivalent non-modified fatty acid controls. Here we demonstrate that laurate (12C), palmitate (16C) and stearate (18C) could activate GDP-inhibited uncoupling protein 1-dependent oxygen consumption in brown adipose tissue mitochondria, whereas there was no activation with glucose-O-ω-laurate (12C), glucose-O-ω-palmitate (16C), glucose-O-ω-stearate (18C), glucose-O-ω-arachidate (20C) or arachidate alone. We conclude that non-flippable fatty acids cannot activate uncoupling protein 1 irrespective of chain length. Our data further undermine the cofactor activation model of uncoupling protein 1 function but are compatible with the model that uncoupling protein 1 functions by flipping long-chain fatty acid anions.

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Abbreviations

BAIB:

[bis(acetoxy)iodo]benzene

BAT:

Brown adipose tissue

DEPT:

Distortionless enhancement by polarization transfer

HMBC:

Heteronuclear multiple-bond correlation

HMQC:

Heteronuclear multiple-quantum correlation

TEMPO:

2,2,6,6-Tetramethylpiperidinyloxyl

THF:

Tetrahydrofuran

UCP:

Uncoupling protein

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Acknowledgments

The research in this manuscript was funded by a Science Foundation Ireland Principal Investigator grant to RKP and a Trinity Studentship award to KJC. RKP is a member of the MITOFOOD COST action (FA0602). Higher Education Authority Programme for Research Cycle 3 for funding to PVM.

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Correspondence to Richard K. Porter.

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Breen, E.P., Pilgrim, W., Clarke, K.J. et al. Lack of activation of UCP1 in isolated brown adipose tissue mitochondria by glucose-O-ω-modified saturated fatty acids of various chain lengths. J Chem Biol 6, 121–133 (2013). https://doi.org/10.1007/s12154-013-0093-6

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Keywords

  • Mitochondria
  • UCP1
  • Glucose-O-ω-fatty acids
  • BAT