Comparative functional analysis reveals differential nucleotide sensitivity between human and mouse UCP1

Aim Mitochondrial uncoupling protein 1 (UCP1) is a unique protein of brown adipose tissue. Upon activation by free fatty acids, UCP1 facilitates a thermogenic net proton flux across the mitochondrial inner membrane. Non‐complexed purine nucleotides inhibit this fatty acid‐induced activity of UCP1. The most available data have been generated from rodent model systems. In light of its role as a putative pharmacological target for treating metabolic disease, in‐depth analyses of human UCP1 activity, regulation, and structural features are essential. Methods In the present study, we established a doxycycline‐regulated cell model with inducible human or murine UCP1 expression and conducted functional studies using respirometry comparing wild‐type and mutant variants of human UCP1. Results We demonstrate that human and mouse UCP1 exhibit similar specific fatty acid‐induced activity but a different inhibitory potential of purine nucleotides. Mutagenesis of non‐conserved residues in human UCP1 revealed structural components in α‐helix 56 and α‐helix 6 crucial for uncoupling function. Conclusion Comparative studies of human UCP1 with other orthologs can provide new insights into the structure–function relationship for this mitochondrial carrier and will be instrumental in searching for new activators.