Abstract
Beta adrenergic receptors (βARs) mediate physiologic responses to the catecholamines epinephrine and norepinephrine released by the sympathetic nervous system. While the hormone epinephrine binds β1AR and β2AR with similar affinity, the smaller neurotransmitter norepinephrine is approximately tenfold selective for the β1AR. To understand the structural basis for this physiologically important selectivity, we solved the crystal structures of the human β1AR bound to an antagonist carazolol and different agonists including norepinephrine, epinephrine and BI-167107. Structural comparison revealed that the catecholamine-binding pockets are identical between β1AR and β2AR, but the extracellular vestibules have different shapes and electrostatic properties. Metadynamics simulations and mutagenesis studies revealed that these differences influence the path norepinephrine takes to the orthosteric pocket and contribute to the different association rates and thus different affinities.
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Data availability
The coordinates and structures factors of T4L–β1AR/carazolol, T4L–β1AR/Nb6B9/BI-167107, T4L–β1AR/Nb6B9/norepinephrine and T4L–β1AR/Nb6B9/epinephrine structures have been deposited in Protein Data Bank under accession number 7BVQ, 7BU7, 7BU6 and 7BTS, respectively.
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Acknowledgements
We gratefully acknowledge the compute resources and support provided by the Erlangen Regional Computing Center (RRZE) and support provided by Radioisotope Laboratory, Center of Biomedical Analysis, Tsinghua University. This work was supported by the Beijing Advanced Innovation Center for Structural Biology, Tsinghua University (X.X. and X.L.), by the DFG grant GRK 1910 (P.G. and J.K.), National Institute of General Medical Sciences GM106990 (B.K.K., P.G. and R.K.S.) and GM083118 (B.K.K. and R.K.S). B.K.K. is a Chan Zuckerberg Biohub investigator and an Einstein BIH Visiting Fellow.
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X.X. performed β1AR expression, purification and crystallization. X.X. and X.L. performed structure determination and refinement. J.K. performed MD simulations, charge and MEP calculations supervised by P.G. X.X., X.L., H.H. and M.J.C. characterized the pharmacology properties of βARs and mutants. M.J.C. performed the binding kinetics assays supervised by R.K.S. K.H. performed automatic data collection and processing. The paper was written by B.K.K. and X.L., with input from X.X. and J.K., and editing and suggestions from P.G. and R.K.S. B.K.K. coordinated the experiments and supervised the overall research. All authors contributed to the editing of the paper.
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B.K.K. is a co-founder of and consultant for ConfometRx, Inc. The other authors declare no competing financial interests.
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Xu, X., Kaindl, J., Clark, M.J. et al. Binding pathway determines norepinephrine selectivity for the human β1AR over β2AR. Cell Res 31, 569–579 (2021). https://doi.org/10.1038/s41422-020-00424-2
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DOI: https://doi.org/10.1038/s41422-020-00424-2
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