Research Paper on The Class D G protein-coupled Yeast -Factor Receptor’s structure and function have been determined experimentally.

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Title: The Class D G protein-coupled Yeast -Factor Receptor’s structure and function have been determined experimentally.

Abstract: The large family of G protein-coupled receptors, which has important medical applications, has been studied using the Saccharomyces cerevisiae -factor mating pheromone receptor (Ste2p) as a model. Many loss-of-function, constitutively active, dominant-negative, and intragenic second-site suppressor mutants as well as mutations that specifically affect pheromone binding were discovered through diverse yeast genetic screens and high-throughput STE2 mutagenesis. The straightforward genetic manipulation of Ste2p also facilitated targeted biochemical methods, such as the use of cysteine disulfide crosslinking to pinpoint intramolecular contacts in the transmembrane helix bundle of Ste2p and contacts between the monomers in a Step2 p dimer as well as probing the aqueous accessibility of substituted cysteine residues to pinpoint the boundaries of the seven transmembrane segments. In contrast to three-dimensional structures exhibiting activation-related conformational changes, recent publication of a series of high-resolution cryo-EM structures of Ste2p in ligand-free, agonist-bound, and antagonist-bound states now makes it possible to evaluate the outcomes of these genetic and biochemical strategies. The outcomes show that the genetic and biochemical approaches were generally successful and offer recommendations for how to best apply these experimental approaches to other proteins. In addition to suggesting aspects of receptor function that go beyond what can be inferred from the available structures, these strategies continue to be helpful in defining mechanisms of signal transduction in the context of the existing structures.

Keywords: protein-coupled receptor; yeast pheromone receptor; yeast mating pathway; substituted cysteine accessibility method; disulfide crosslinking; second-site suppressor; constitutively active receptor; dominant-negative receptor; receptor oligomerization

Paper Quality: SCOPUS / Web of Science Level Research Paper

Subject: Biomolecules

Writer Experience: 20+ Years

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