Analysis of parathyroid hormone's principal receptor-binding region by site-directed mutagenesis and analog design.

Gardella, Thomas J.; Wilson, Andrew; Keutmann, Henry T.; Oberstein, Robert; Potts, John T.; Kronenberg, Mark S.; Nussbaum, Samuel R.

doi:10.1210/endo.132.5.8386605

Cited by 60 publications

(55 citation statements)

References 33 publications

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“…In contrast, other mutationally intolerant ligand residues nearby (e.g. at positions 23, 28, and 31 (16,17)) tolerated the Bpa substitution with no apparent reduction in binding affinity. At position 23, Bpa replaced a tryptophan, and the structural similarity between these two residues may explain why the substitution was tolerated.…”

Section: Analogsmentioning

confidence: 88%

“…Mapping (17). LysC digestion of the covalent complex obtained with this ligand revealed two overlapping nonglycosylated fragments of 4 -5 kDa (Fig.…”

Section: Analogsmentioning

confidence: 99%

“…Binding to the P1R was only minimally affected by most of the Bpa substitutions, with the exception of [I 5 ,W 23 , Bpa 24 ,Y 36 ]PTHrP-(1-36)-amide, which showed an ϳ10-fold reduced ability to compete with 125 I-labeled rPTH-(1-34) at the hP1R (Table I). The conserved Leu at position 24 in PTH was previously shown to be intolerant to substitution with polar residues (ϳ1000-fold reduced binding affinity) (17). In contrast, other mutationally intolerant ligand residues nearby (e.g.…”

Section: Analogsmentioning

confidence: 99%

“…Two residues within this region, Thr 33 and Gln 37 , were shown to be important for binding of PTHrP- (18). Residues 24 -34 of PTHrP-(1-34) (19) and PTH-(1-34) (20) also contribute to high affinity binding to the P1R, and three hydrophobic amino acids within this region of PTH-(1-34), residues Leu 24 , Leu 28 , and Val 31 , have been found to be intolerant to substitution with polar residues (17). A PTH analog with a benzoyl group attached to the epsilonamino group of lysine 27 has been shown to cross-link to extracellular loop 1 of the human P1R (21), a result that would not have been predicted by the previous receptor mutagenesis and chimera studies (3,8,13).…”

mentioning

confidence: 99%

“…1 below). Ligand residue 23 (Phe in PTHrP and Trp in PTH) has been found to be important for high affinity binding to the P1R and to the PTH-2 receptor (16,17). Replacement of this residue with a Bpa photoreactive group results in affinity cross-linking to residues 23-40 of the rat P1R, at the extreme end of the amino-terminal extracellular domain.…”

mentioning

confidence: 99%

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Multiple Sites of Contact between the Carboxyl-terminal Binding Domain of PTHrP-(1–36) Analogs and the Amino-terminal Extracellular Domain of the PTH/PTHrP Receptor Identified by Photoaffinity Cross-linking

Gensure¹,

Gardella²,

Jüppner³

2001

Journal of Biological Chemistry

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The carboxyl-terminal portions of parathyroid hormone (PTH)-(1-34) and PTH-related peptide (PTHrP)-(1-36) are critical for high affinity binding to the PTH/ PTHrP receptor (P1R), but the mechanism of receptor interaction for this domain is largely unknown. To identify interaction sites between the carboxyl-terminal region of PTHrP-(1-36) and the P1R, we prepared analogs of [ -172. These data thus predict that residues 23, 27, 28, and 33 of native PTHrP are each near to different regions of the amino-terminal extracellular receptor domain of the P1R. This information helps define sites of proximity between several ligand residues and this large receptor domain, which so far has been largely excluded from models of the hormone-receptor complex. PTH and PTHrP1 mediate many of their biological effects through the same receptor (1, 2). Peptides containing the first 34 amino acids of PTH and PTHrP are capable of fully activating the PTH/PTHrP receptor (P1R) (1, 2). Studies with PTH and PTHrP ligand analogs and receptor chimeras have suggested that the ligands have two distinct functional domains: the amino-terminal residues, which are important for receptor activation; and the carboxyl-terminal residues, which are important for high affinity binding (3). These data furthermore indicate that the ligand's amino-terminal portion interacts with the extracellular loops and the membrane-spanning helices of the receptor, and the ligand's carboxyl-terminal portion interacts with the receptor's amino-terminal extracellular domain (3). A similar pattern of ligand-receptor interaction has been suggested for other members of this class II family of peptide hormone G-protein-coupled receptors, including the secretin receptor (4) and the PTH-2 receptor (5-7).Some specific sites of interaction between the amino-terminal portions of PTH-(1-34)/PTHrP-(1-36) and the P1R have been identified by site-directed mutagenesis and photoaffinity cross-linking studies. For example, mutational analyses have shown that residues in extracellular loop 3 and the adjacent sixth membrane-spanning helix (TM6) are critical for mediating ligand-induced receptor activation (2,8). Consistent with these mutational data, Bpa introduced at position 1 of PTH-(1-34) or position 2 of either PTH-(1-34) or PTHrP-(1-36) was found to cross-link to methionine 425, at the extracellular end of TM6 in the P1R (9, 10). Interactions between the aminoterminal portion of the ligand and the transmembrane domains/extracellular loops of the receptor are also suggested by a study showing that PTH-(1-14) can stimulate cAMP accumulation in a mutant P1R missing most of the amino-terminal extracellular domain as efficiently as it does in the wild-type P1R (11). Furthermore, if a PTH fragment comprising the first 9 amino acids is covalently attached to the amino-terminal end of such a truncated receptor, the resulting ligand-receptor chimera displays constitutive activity, indicative of an intramolecular stimulation of the receptor's activation domain by the tethered ligand fragment ...

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Section: Analogsmentioning

confidence: 88%

“…Mapping (17). LysC digestion of the covalent complex obtained with this ligand revealed two overlapping nonglycosylated fragments of 4 -5 kDa (Fig.…”

Section: Analogsmentioning

confidence: 99%

Section: Analogsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Multiple Sites of Contact between the Carboxyl-terminal Binding Domain of PTHrP-(1–36) Analogs and the Amino-terminal Extracellular Domain of the PTH/PTHrP Receptor Identified by Photoaffinity Cross-linking

Gensure¹,

Gardella²,

Jüppner³

2001

Journal of Biological Chemistry

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Peptide and Protein Hormones, Peptide Neurotransmitters, and Therapeutic Agents

Hruby

Chavez

Kavarana

2003

Burger's Medicinal Chemistry and Drug Discovery

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Over 50% of all current drugs and perhaps an even higher percentage of drug targets have as their major target receptors/acceptors for which the native ligand/recognition motif is a peptide. Furthermore, most targets for drugs are proteins or conjugated proteins. Peptide hormones and neurotransmitters constitute a very large number of these ligands and almost all of them are important in a wide variety of diseases. Thus robust methods for the design, synthesis, and biological evaluation of peptides, proteins, peptide analogs, and peptidomimetics are needed. In this chapter we briefly discuss the major synthetic, structural, conformational, and topographical considerations that go into peptide design of peptide analogs, derivatives, and mimetics that can serve as drug candidates. We then provide a selected discussion on the design of four different examples of peptide hormone and neurotransmitter ligands, and conclude with a brief discussion of future perspectives in this critically important area of current and future drug research.

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PTH and Vitamin D

Khundmiri

Murray

Lederer

2016

Comprehensive Physiology

234

158

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PTH and Vitamin D are two major regulators of mineral metabolism. They play critical roles in the maintenance of calcium and phosphate homeostasis as well as the development and maintenance of bone health. PTH and Vitamin D form a tightly controlled feedback cycle, PTH being a major stimulator of vitamin D synthesis in the kidney while vitamin D exerts negative feedback on PTH secretion. The major function of PTH and major physiologic regulator is circulating ionized calcium. The effects of PTH on gut, kidney, and bone serve to maintain serum calcium within a tight range. PTH has a reciprocal effect on phosphate metabolism. In contrast, vitamin D has a stimulatory effect on both calcium and phosphate homeostasis, playing a key role in providing adequate mineral for normal bone formation. Both hormones act in concert with the more recently discovered FGF23 and klotho, hormones involved predominantly in phosphate metabolism, which also participate in this closely knit feedback circuit. Of great interest are recent studies demonstrating effects of both PTH and vitamin D on the cardiovascular system. Hyperparathyroidism and vitamin D deficiency have been implicated in a variety of cardiovascular disorders including hypertension, atherosclerosis, vascular calcification, and kidney failure. Both hormones have direct effects on the endothelium, heart, and other vascular structures. How these effects of PTH and vitamin D interface with the regulation of bone formation are the subject of intense investigation.

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Analysis of parathyroid hormone's principal receptor-binding region by site-directed mutagenesis and analog design.

Cited by 60 publications

References 33 publications

Multiple Sites of Contact between the Carboxyl-terminal Binding Domain of PTHrP-(1–36) Analogs and the Amino-terminal Extracellular Domain of the PTH/PTHrP Receptor Identified by Photoaffinity Cross-linking

Multiple Sites of Contact between the Carboxyl-terminal Binding Domain of PTHrP-(1–36) Analogs and the Amino-terminal Extracellular Domain of the PTH/PTHrP Receptor Identified by Photoaffinity Cross-linking

Peptide and Protein Hormones, Peptide Neurotransmitters, and Therapeutic Agents

PTH and Vitamin D

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