The Unpredicted High Affinities of a Large Number of Naturally Occurring Tachykinins for Chimeric NK1/NK3 Receptors Suggest a Role for an Inhibitory Domain in Determining Receptor Specificity

Tian, Ye; Wu, Lan-Hsin; Oxender, Dale L.; Chung, Fung Lung

doi:10.1074/jbc.271.34.20250

Cited by 27 publications

(14 citation statements)

References 26 publications

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“…48 Using chimeric NK 1 and NK 3 receptors, Tian et al have shown that the C-terminal sequence of SP is important in determining binding affinities, while the N-terminal sequence seems to be more important in maintaining the appropriate conformation of the peptide for receptor binding and selectivity. 49 In this study we have shown that residues 4-9 of RTKA, RTKB, and RTKC exist in a similar conformation as determined in a SDS micelle system. This findings as well as the amino acid conservation supports the hypothesis that the biological activity and receptor binding to the bfSPR are confined to the C-terminus.…”

Section: Discussionmentioning

confidence: 71%

Solution Structures in SDS Micelles and Functional Activity at the Bullfrog Substance P Receptor of Ranatachykinin Peptides

et al. 2000

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A set of novel tachykinin-like peptides has been isolated from bullfrog brain and gut. These compounds, ranatachykinin A (RTKA), ranatachykinin B (RTKB), and ranatachykinin C (RTKC), were named for their source, Rana catesbeiana, and their homology to the tachykinin peptide family. We present the first report of the micelle-bound structures and pharmacological actions of the RTKs. Generation of three-dimensional structures of the RTKs in a membrane-model environment using (1)H NMR chemical shift assignments, two-dimensional NMR techniques, and molecular dynamics and simulated annealing procedures allowed for the determination of possible prebinding ligand conformations. RTKA, RTKB, and RTKC were determined to be helical from the midregion to the C-terminus (residues 4-10), with a large degree of flexibility in the N-terminus and minor dynamic fraying at the end of the C-terminus. The pharmacological effects of the RTKs were studied by measuring the elevation of intracellular Ca(2+) in Chinese hamster ovarian cells stably transfected with the bullfrog substance P receptor (bfSPR). All of the RTKs tested elicited Ca(2+) elevations with a rank order of maximal effect of RTKA >/= SP > RTKC >/= RTKB. A high concentration (1 microM) of the neuropeptides produced varying degrees of desensitization to a subsequent challenge with the same or different peptide, while a low concentration (1 pM) produced sensitization at the bfSPR. Our data suggest differences in amino acid side chains and their charged states at the C-terminal sequence or differences in secondary structure at the N-terminus, which do not overlap according to the findings in this paper, may explain the differing degree and type of receptor activation seen at the bfSPR.

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Section: Discussionmentioning

confidence: 71%

Solution Structures in SDS Micelles and Functional Activity at the Bullfrog Substance P Receptor of Ranatachykinin Peptides

et al. 2000

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“…This diversification appears to have evolved by the emergence of inhibitory domains on both ligand and receptor which impose steric hindrances thus allowing only the intended ligand to interact with the common activation domain (28). Such negative specificity determinants have not only developed in glycoprotein hormones and their receptors, but also in other members of the cystine knot growth factor superfamily, such as neurotropins (29), and also in other G protein-coupled receptors (30). More generally, dimer formation is necessary for the activity and specificity of many, if not all cystine knot growth factors, as well as for other bioactive molecules, such as enzymes and transcription factors.…”

Section: Discussionmentioning

confidence: 99%

Human Thyroid-stimulating Hormone (hTSH) Subunit Gene Fusion Produces hTSH with Increased Stability and Serum Half-life and Compensates for Mutagenesis-induced Defects in Subunit Association

Grossmann

Wong²,

Szkudlinski

et al. 1997

Journal of Biological Chemistry

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The human thyroid-stimulating hormone (hTSH) subunits ␣ and ␤ are transcribed from different genes and associate noncovalently to form the bioactive hTSH heterodimer. Dimerization is rate-limiting for hTSH secretion, and dissociation leads to hormone inactivation. Previous studies on human chorionic gonadotropin (hCG) and human follicle-stimulating hormone had shown that it was possible by subunit gene fusion to produce a bioactive, single chain hormone. However, neither the stability nor the clearance from the circulation of such fused glycoprotein hormones has been studied. We show here that genetic fusion of the hTSH ␣-and ␤-subunits using the carboxyl-terminal peptide of the hCG ␤-subunit as a linker created unimolecular hTSH whose receptor binding and bioactivity were comparable to native hTSH. Interestingly, the fused hTSH had higher thermostability and a longer plasma half-life than either native or dimeric hTSH containing the hCG ␤-subunit-carboxyl-terminal peptide, suggesting that dimer dissociation may contribute to glycoprotein hormone inactivation in vivo. In addition, we show for the first time that synthesis of hTSH as a single polypeptide chain could overcome certain mutagenesis-induced defects in hTSH secretion, therefore enabling functional studies of such mutants. Thus, in addition to prolongation of plasma half-life, genetic fusion of hTSH subunits should be particularly relevant for the engineering of novel analogs where desirable features are offset by decreased dimer formation or stability. Such methods provide a general approach to expand the spectrum of novel recombinant glycoprotein hormones available for in vitro and in vivo study.

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“…In addition, an attempt to determine the basis of multiallelic self-incompatibility in plants was carried out by exchanging domains between allelic S-RNases from Nicotiana alata (39). Chimeric proteins have also been used to study proteinprotein interactions during ligand-receptor recognition (7,20,21,32). In fact, our observation that bE and bW class II chimeric alleles have specificities different from either parent is not unique to fungal mating-type systems; a similar phenomenon has recently been reported for chimeras of two glycoprotein hormones (4).…”

Section: Figmentioning

confidence: 99%

Dual Sets of Chimeric Alleles Identify Specificity Sequences for the bE and bW Mating and Pathogenicity Genes of Ustilago maydis

Yee

Kronstad

1998

Molecular and Cellular Biology

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The b mating-type locus of the fungal plant pathogen Ustilago maydis encodes two multiallelic gene products, bE and bW, that control the formation and maintenance of the infectious cell type. Dimerization via the N-terminal regions of bE and bW proteins encoded by alleles of different specificities establishes a homeodomain-containing transcription factor. The bE and bW products encoded by alleles of like specificities fail to dimerize. We constructed sets of chimeric alleles for the bE1 and bE2 genes and for the bW1 and bW2 genes to identify sequences that control specificity. The mating behavior of strains carrying chimeric alleles identified three classes of specificity: b2 (class I), specificity different from either parental type (class II), and b1 (class III). Crosses between strains carrying bE and bW chimeric alleles identified two short blocks of amino acids that influence specificity and that are located in the N-terminal variable regions of the b proteins. Comparisons of pairs of chimeric alleles encoding polypeptides differing in specificity and differing at single amino acid positions identified 16 codon positions that influence the interaction between bE and bW. Fifteen of these positions lie within the blocks of amino acids identified by crosses between the strains carrying chimeric alleles. Overall, this work provides insight into the organization of the regions that control recognition.Recognition mediated by protein-protein interactions plays a fundamental role in many biological processes. Well-characterized examples include antibody-antigen interactions (8, 9, 23), ligand-receptor binding (22,35), and the establishment and maintenance of tissue integrity by cadherins (19). The proteins involved in sexual reproduction and incompatibility in fungi provide relatively simple examples of determinants of self versus nonself recognition. In this paper, we describe a molecular genetic approach to identify the determinants of recognition for the proteins encoded by the b mating-type locus of the fungal corn pathogen Ustilago maydis.U. maydis is commonly found in nature as black diploid teliospores on infected corn plants (6). The teliospores germinate, and meiosis occurs to produce haploid, yeast-like progeny. Nonself recognition between compatible haploid mating partners is a prerequisite to the establishment of an infectious, dikaryotic cell type, and the genes at the a and b mating-type loci are considered pathogenicity factors (reviewed in references 2 and 18). The a locus, with alternate specificities a1 and a2, encodes pheromones and pheromone receptors and controls recognition of mating partners at the level of cell fusion (3,11,31). The b locus controls the formation and maintenance of the infectious cell type after cell fusion has occurred. If the cells participating in mating have different specificities (nonself) at the b locus, a vigorous, straight dikaryotic filament is formed and this cell type will be infectious. In contrast, mating partners that carry b sequences of like specificities ...

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The Unpredicted High Affinities of a Large Number of Naturally Occurring Tachykinins for Chimeric NK1/NK3 Receptors Suggest a Role for an Inhibitory Domain in Determining Receptor Specificity

Cited by 27 publications

References 26 publications

Solution Structures in SDS Micelles and Functional Activity at the Bullfrog Substance P Receptor of Ranatachykinin Peptides

Solution Structures in SDS Micelles and Functional Activity at the Bullfrog Substance P Receptor of Ranatachykinin Peptides

Human Thyroid-stimulating Hormone (hTSH) Subunit Gene Fusion Produces hTSH with Increased Stability and Serum Half-life and Compensates for Mutagenesis-induced Defects in Subunit Association

Dual Sets of Chimeric Alleles Identify Specificity Sequences for the bE and bW Mating and Pathogenicity Genes of Ustilago maydis

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