Identification of a Contact Region between the Tridecapeptide α-Factor Mating Pheromone of<i>Saccharomyces cerevisiae</i>and Its G Protein-Coupled Receptor by Photoaffinity Labeling

Henry, L. Keith; Khare, Sanjay K.; Son, Çağdaş Devrim; Babu, Vommina V. Suresh; Naider, Fred; Becker, Jeffrey M.

doi:10.1021/bi015863z

Cited by 40 publications

(62 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…16,17 Among the many synthetic β-turn peptidomimetics restricted by cyclization, (R)-γ-lactam conformational constraint incorporating [3-(R)-amino-2-oxo-1-pyrrolidineacetamido] in place of ProGly at residues 8 and 9 was reported to have the best activity only equal to that of parent peptide, [Nle ing approach involving the photoactivatable groups attached to the α-factor backbone was developed. 28,30,44,[51][52][53]55,56 pBenzoyl-L-phenylalanine (Bpa) and 3,4-Dihydroxyphenylalanine (DOPA) was reported to have a desirable property for this purpose, although incorporation of these groups at various positions in α-factor have been shown to result in a less than 30-fold decrease in receptor affinity. 44,51,55 Biotin (Bio) tagging modification of DOPA analog (Bio 7…”

Section: 16mentioning

confidence: 99%

“…28,30,44,[51][52][53]55,56 pBenzoyl-L-phenylalanine (Bpa) and 3,4-Dihydroxyphenylalanine (DOPA) was reported to have a desirable property for this purpose, although incorporation of these groups at various positions in α-factor have been shown to result in a less than 30-fold decrease in receptor affinity. 44,51,55 Biotin (Bio) tagging modification of DOPA analog (Bio 7…”

Section: 16mentioning

confidence: 99%

“…Although hundreds of α-factor analogs have been reported over the last thirty years, 4,42,43,51,53 most of these synthetic analogs are known to markedly reduce activity, and only a few have activities comparable with that of the native α-factor. Surprisingly, none of these analogs exceed higher than 3-to 4-fold activity compared to that of native α-factor.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Highly Active Analogs of α-Factor and Their Activities Against Saccharomyces cerevisiae

Ahn¹,

Hong²,

Jin³

et al. 2014

Bulletin of the Korean Chemical Society

View full text Add to dashboard Cite

Thirteen analogs of tridecapeptide α-factor (WHWLQLKPGQPMY) of Saccharomyces cerevisiae with C-or N-terminal Trp extension and isosteric replacement by Aib at position 8 and 11, Trp at position 13, D-Ala at position 9, and Orn and Glu at position 6 were synthesized and assayed for their biological activity. Receptor binding assay was carried out using our newly developed spectrophotometric method with detector peptide 14. C-or N-terminal extended analogs, α-factor-[Trp] n (n =1-5) 1-5 and [N-Trp] 1 -α-factor 6, were all less active than native α-factor and gradual decreases in both activity and receptor affinity were observed with greater Trp extension. Trp-substituted analog at position 13, [Trp 13 ]α-factor 7, exhibited about 2-fold reductions in both activity and receptor affinity. Aib-substituted analogs, [Aib 8 ]α-factor 8 and [Aib 11]α-factor 9, showed 5-to 10-fold reduction in activity as well as 3-fold reduction in receptor affinity compared to native α-factor. [Orn 6]α-factor 10 demonstrated strong potency with a 7.0-fold increase in halo activity as well as 1.8-fold increase in receptor affinity compared to native α-factor. ]α-factor 11 exhibited 15-fold higher halo activity as well as nearly 3-fold higher receptor affinity compared to native α-factor.

show abstract

Section: 16mentioning

confidence: 99%

Section: 16mentioning

confidence: 99%

See 1 more Smart Citation

Highly Active Analogs of α-Factor and Their Activities Against Saccharomyces cerevisiae

Ahn¹,

Hong²,

Jin³

et al. 2014

Bulletin of the Korean Chemical Society

View full text Add to dashboard Cite

show abstract

“…As with other GPCR proteins and in keeping with the crystal structure of rhodopsin (31), there are seven TM domains, and the third TM domain appears unusually long. The N terminus is predicted to be extracellular, and the intracellular loops TM3-4 and TM5-6 are thought to form a cleft in which the cognate G protein ␣-subunit binds (32)(33)(34). There is a clear relationship between codon diversity and TM topology of SRZ proteins.…”

Section: Positive Selection In the Sr Superfamilymentioning

confidence: 99%

Adaptive evolution in the SRZ chemoreceptor families of Caenorhabditis elegans and Caenorhabditis briggsae

Thomas

Kelley

Robertson

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

We investigated the possibility of positive selection acting on members of the putative seven-pass chemoreceptor superfamily in Caenorhabditis elegans, which comprises Ϸ1,300 genes encoding seven-pass G protein-coupled receptors (GPCRs). Using a maximum-likelihood approach, we conducted statistical tests for evidence of codon sites where the ratio of nonsynonymous substitutions per nonsynonymous site to synonymous substitutions per synonymous site (d N͞dS) was >1. Evidence for positive selection was found only for the srz family, about which virtually nothing specific is known. We extended the annotation of the srz gene family, establishing gene models for 60 srz genes in C. elegans and 28 srz genes in Caenorhabditis briggsae. d N͞dS ratios varied dramatically in different regions of the SRZ proteins, peaking in predicted extracellular regions. These regions included 23 sites where evidence of positive selection was highly significant, corresponding remarkably well with regions implicated in ligand binding in other GPCR family members. We interpret these results as indicating that the srz family is under positive selection, probably driven by ligand binding.positive selection ͉ ligand binding ͉ maximum likelihood ͉ synonymous ͉ nonsynonymous C aenorhabditis elegans has Ϸ1,300 predicted genes that encode members of putative chemosensory receptors and together define the seven-pass receptor (SR) superfamily (1-4), which belongs to the broader class of G protein-coupled receptors (GPCRs). Based on sequence alignment and phylogenetic analysis, SR superfamily members fall into about a dozen families. These families range in size from the large srh and str families (a few hundred genes each) to the modestly sized sra and srv families (Ϸ30 genes each). Each SR family appears to have arisen by gene duplication and divergence from a founder gene. These duplications have occurred sporadically over a long evolutionary period, giving rise to complex relationships. Near one extreme, str-5 and str-6 result from a recent duplication and differ by only two nucleotides in their coding regions. Near the other extreme, the str-1 and str-47 proteins are only 19% identical and presumably arose from an ancient duplication. Members of different SR families are even more distantly related, with the most distant pairs barely alignable.Proteins in the SR superfamily appear to be more rapidly diverging than the average gene (2, 3, 5, 6). For example, the average ortholog pair between C. elegans and Caenorhabditis briggsae has 80% amino acid identity (5), whereas the average str pair from the same data set has 59% identity (J.H.T., unpublished data). Rapidly diverging proteins may result from relaxed selective constraints, in which changes in protein sequence are relatively well tolerated. Alternatively, they may result from selective pressure for changes in amino acid sequence (positive selection or diversifying selection). In extreme cases, positive selection acts on all or most of a protein and can be detected in pairwise d N ͞d S ratio...

show abstract

“…Coupling energies found for two directly interacting residues can be converted to a distance constraint and in this respect are similar in nature to nuclear Overhauser connectivities (11). Thus, detailed knowledge of coupling energies for a ligand and a receptor can be used to dock the ligand into a receptor whose structure is available (12-14).We have been studying the biology of a yeast mating factor GPCR, Ste2p, and its contacts with α-factor [Trp 1 -His-Trp-Leu-Gln-Leu-Lys-Pro-Gly-Gln-Pro-Met-Tyr 13 ], the tridecapeptide ligand of this GPCR, by employing α-factor analogs, site-specific mutagenesis and photocrosslinking analysis (15)(16)(17)(18). These studies together with extensive molecular biology investigations (19-23) have indicated likely interactions between Y266 and residues near the amine terminus of α-factor.…”

mentioning

confidence: 99%

Double-Mutant Cycle Scanning of the Interaction of a Peptide Ligand and Its G Protein-Coupled Receptor

et al. 2007

Self Cite

View full text Add to dashboard Cite

The interaction between the yeast G protein coupled receptor (GPCR), Ste2p, and its α-factor tridecapeptide ligand was subjected to double-mutant cycle scanning analysis by which the pairwise interaction energy of each ligand residue with two receptor residues, N205 and Y266, was determined. The mutations N205A and Y266A were previously shown to result in deficient signaling but cause only a 2.5-fold and 6-fold decrease, respectively, in the affinity for α-factor. The analysis shows that residues at the amine terminus of α-factor interact strongly with N205 and Y266 whereas residues in the center and at the carboxyl terminus of the peptide interact only weakly if at all with these receptor residues. Multiple-mutant thermodynamic cycle analysis was used to assess whether the energies of selected pairwise interactions between residues of the α-factor peptide changed upon binding to Ste2p. Strong positive cooperativity between residues 1 through 4 of α-factor was observed during receptor binding. In contrast, no thermodynamic evidence was found for an interaction between a residue near the carboxyl terminus of α-factor (position 11) and one at the N-terminus (position 3). The study shows that multiple-mutant cycle analyses of the binding of an alaninescanned peptide to wild-type and mutant GPCRs can provide detailed information on contributions of inter-and intra-molecular interactions to the binding energy and potentially prove useful in developing 3D models of ligand docked to its receptor.Determination of contacts between peptide ligands and their cognate G protein-coupled receptors (GPCRs) 2 and the respective energetics of these interactions is essential for understanding how binding is transduced to intracellular signaling. Many studies have employed structure-activity relationships involving ligand analogs and mutagenesis of receptors to discern receptor-ligand interactions (reviewed in refs.1,2). Complementary investigations have used photocrosslinking to provide biochemical evidence for contacts (3,4). In the 1980s, Fersht and coworkers and Horovitz introduced double-mutant cycle analysis to provide thermodynamic evidence for the interaction between groups within one protein or in ligand-protein complexes (5-7). The method has been applied to a number of receptor systems including the ligand-gated ion channel nicotinic (8,9) and the GPCR muscarinic (10) acetylcholine receptors. The concept of this method is illustrated in a cycle for a ligand binding to its GPCR ( Figure 1A). If the effect on the binding free energy (or the free energy of some other process) of the double mutation is not equal to the sum of effects of the single mutations then the two residues are coupled. Non-zero pairwise coupling energies calculated from such † This work was supported by research grants GM22086 and GM22087 from the National Institutes of Health. cycles reflect interactions that can be either direct or indirect. Coupling energies found for two directly interacting residues can be converted to a distance constrain...

show abstract

Identification of a Contact Region between the Tridecapeptide α-Factor Mating Pheromone ofSaccharomyces cerevisiaeand Its G Protein-Coupled Receptor by Photoaffinity Labeling

Cited by 40 publications

References 31 publications

Highly Active Analogs of α-Factor and Their Activities Against Saccharomyces cerevisiae

Highly Active Analogs of α-Factor and Their Activities Against Saccharomyces cerevisiae

Adaptive evolution in the SRZ chemoreceptor families of Caenorhabditis elegans and Caenorhabditis briggsae

Double-Mutant Cycle Scanning of the Interaction of a Peptide Ligand and Its G Protein-Coupled Receptor

Contact Info

Product

Resources

About