Structure of the complement factor 5a receptor-ligand complex studied by disulfide trapping and molecular modeling

The Journal of Biological Chemistry
Ian S HagemannThomas J Baranski

Abstract

Complement factor 5a (C5a) is an anaphylatoxin that acts by binding to a G protein-coupled receptor, the C5aR. The relative orientation of this ligand-receptor pair is investigated here using the novel technique of disulfide trapping by random mutagenesis (DTRM) and molecular modeling. In the DTRM technique, an unpaired cysteine is introduced in the ligand, and a library of randomly mutagenized receptors is screened to identify mutants that introduce a cysteine at a position in the receptor that allows functional interactions with the ligand. By repeating this analysis at six positions of C5a, we identify six unique sets of intermolecular interactions for the C5a-C5aR complex, which are then compared with an independently developed computational three-dimensional model of the complex. This analysis reveals that the interface of the receptor N terminus with the cysteine-containing ligand molecules is selected from a variety of possible receptor conformations that exist in dynamic equilibrium. In contrast, DTRM identifies a single position in the second extracellular loop of the receptor that interacts specifically with a cysteine probe placed in the C-terminal tail of the C5a ligand.

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Citations

Jun 10, 2008·Nature Immunology·Folmer FredslundGregers R Andersen
Sep 12, 2012·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Maoqing DongLaurence J Miller
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Oct 6, 2009·Biochemical and Biophysical Research Communications·Gregory V Nikiforovich, Thomas J Baranski
Jul 16, 2016·Biochemical and Biophysical Research Communications·Da-Eun HwangHak-Sung Kim
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Oct 21, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Anette Kaiser, Irene Coin
Aug 27, 2019·Bioorganic & Medicinal Chemistry·Richa Mishra, Soumendra Rana
Feb 16, 2008·Biochemistry·Gregory V NikiforovichThomas J Baranski

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