Analogues of parathyroid hormone modified at positions 3 and 6. Effects on receptor binding and activation of adenylyl cyclase in kidney and bone

Cohen, Fred E.; Strewler, Gordon J; Bradley, Mary S.; Carlquist, Mats; Nilsson, Magnus; Ericsson, Maja; Ciardelli, Thomas L.; Nissenson, Robert A.

doi:10.1016/s0021-9258(18)52390-5

Cited by 75 publications

(10 citation statements)

References 52 publications

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“…) -4500 deg cm 2 dmol -1 , 22 compared to an average value of -6800 deg cm 2 dmol -1 reported for hPTH under similar conditions). 20,[23][24][25][26] Of course, an increase in R-helix content is observed upon addition of TFE, producing CD spectra almost identical to the corresponding data for PTH . 22 NMR studies on [A 26 ]hPTHrP localize the helix at the N-and C-termini.…”

Section: Introductionsupporting

confidence: 58%

“…Nevertheless, the helix is wellcharacterized by the combination of NOEs, temperature coefficients, and chemical shifts. The beginning of the helix was determined by NOEs from the H R and H β of D 17 to the HN of R 20 . HN(i)-HN(i+1) correlations were observed from residue 17 to 34 (except for NOEs 23/24 and 26/27 that could not be detected owing to the small chemical shift difference between these two pairs of protons).…”

Section: Resultsmentioning

confidence: 99%

“…A closer inspection of the amino acid sequence of RS-66271 reveals that many important features of the parent peptide are maintained. For example, the basic triplet R 19 -R 20 -R 21 preceding the MAP fragment and residues L 24 and L 27 are conserved (see Chart 1). In addition, the substitution at positions 28 and 31 (from isoleucine in hPTHrP to leucine in RS-66271) is conservative, while E 22 ), the binding affinity is restored.…”

Section: Discussionmentioning

confidence: 99%

“…For the PTHrP(1−34) sequence, the aqueous solution conformation is predominantly unstructured, as indicated by the low value of ellipticity at 222 nm in CD spectra ([ϑ r ] 222 = −4500 deg cm 2 dmol -1 , compared to an average value of −6800 deg cm 2 dmol -1 reported for hPTH under similar conditions). ,− Of course, an increase in α-helix content is observed upon addition of TFE, producing CD spectra almost identical to the corresponding data for PTH(1−34) . NMR studies on [A 26 ]hPTHrP(1−34) localize the helix at the N- and C-termini .…”

Section: Introductionmentioning

confidence: 88%

“…The majority of the structural studies by NMR agree that hPTH(1−34) does not form secondary structure elements in nearly physiological solvent conditions, − the only ordered region encompassing residues 20−24 . The sequence-encoded propensity for α-helix is manifest only in a structure-enhancing solvent such as trifluoroethanol, TFE (α-helix in the regions 3−9/12 and 17−26/28 has been reported in different NMR studies). − , …”

Section: Introductionmentioning

confidence: 97%

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Conformational Studies of RS-66271, an Analog of Parathyroid Hormone-Related Protein with Pronounced Bone Anabolic Activity

et al. 1997

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Both the parathyroid hormone (PTH) and the functionally similar parathyroid hormone-related protein (PTHrP) have served as templates for the development of novel bone anabolic agents for the treatment of osteoporosis. The PTHrP analog RS-66271 (Vickery, B. H.; Avnur, Z.; Cheng Y.; Chiou, S.-S.; Leaffer, D.; Caulfield, J. P.; Kimmel, D. B.; Ho, T.; Krstenansky, J. L. J. Bone Miner. Res. 1996, 11, 1943-1951), in which the amino acids 22-31 have been substituted by the sequence E22-L-L-E-K-L-L-E-K-L31 (a model amphiphilic peptide), is a potent bone anabolic agent in vivo. Therefore, RS-66271 is a good candidate for structural analysis with the aim of developing a structure-activity relationship. The structural characterization described here was carried out in aqueous solution employing circular dichroism and nuclear magnetic resonance spectroscopy. We find that the incorporated amphiphilic decapeptide is indeed helical. In addition, it induces the adjacent residues, up to residue 16, to adopt the helical conformation. The helical domain, including residues 16-32, incorporates most of the previously identified principal receptor binding domain PTHrP(25-34). We discuss the relevance of the distinct and extensive helicity in light of the reduced in vitro receptor affinity/ activity and the enhanced in vivo bone anabolic efficacy of RS-66271.

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Section: Introductionsupporting

confidence: 58%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 88%

Section: Introductionmentioning

confidence: 97%

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Conformational Studies of RS-66271, an Analog of Parathyroid Hormone-Related Protein with Pronounced Bone Anabolic Activity

et al. 1997

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Conformational studies of parathyroid hormone (PTH)/PTH-related protein (PTHrP) point-mutated hybrids

et al. 1999

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Conformational studies of parathyroid hormone (PTH)/PTH-related protein (PTHrp) chimeric peptides

et al. 2000

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The N‐terminal 1–34 segments of both parathyroid hormone (PTH) and parathyroid hormone‐related protein (PTHrP) bind and activate the same membrane‐embedded G protein‐coupled receptor (PTH1 Rc) present on the surface of cells in target tissues such as bone and kidney. This binding occurs in spite of major differences between the two hormones in their amino acid sequence. Recently, it was shown that in (1–34) PTH/PTHrP hybrid peptides, the N‐terminal 1–14 segment of PTHrP is incompatible with the C‐terminal 15–34 region of PTH in terms of bioactivity. The sites of incompatibility were identified at positions 5 in PTHrP and 19 in PTH. In the present paper we describe the synthesis, biological evaluation, and conformational characterization of two segmental hybrids: PTHrP(1–27)‐[Tyr34]bPTH(28–34)‐NH2 (hybrid I) and PTHrP(1–18)‐[Nal23, Tyr34]bPTH(19–34)‐NH2 (hybrid II). Hybrid I is as active as PTH(1–34)NH2 and more than two orders of magnitude more active than hybrid II. The conformational properties of the hybrids were studied in water/trifluoroethanol (TFE) mixtures and in aqueous solutions containing dodecylphosphocholine (DPC) micelles by CD, two‐dimensional nmr and computer simulations. Upon addition of TFE to the aqueous solution, both hybrids undergo a coil–helix transition. The helix content in 1:1 water/TFE obtained by CD data is about 75% for both hybrids. In the presence of DPC, helix formation is observed at detergent concentrations above critical micellar concentration and the maximum helix content is of ∼35 and ∼30% for hybrid I and II, respectively. Combined nmr analysis, distance geometry, and molecular dynamics calculations suggest that, in both solvent systems, the biologically active hybrid I exhibits two flexible sites, centered at residues 12 and 19, connecting helical segments. The flexibility point at position 19 is not present in the poorly active hybrid II. Our findings support the hypothesis, proposed in our previous work, that in bioactive PTH analogues the presence and location of flexibility points between helical segments are essential for enabling them to fold into the bioactive conformation upon interaction with the PTH1 receptor. © 2000 John Wiley & Sons, Inc. Biopoly 54: 429–447, 2000

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Analogues of parathyroid hormone modified at positions 3 and 6. Effects on receptor binding and activation of adenylyl cyclase in kidney and bone

Cited by 75 publications

References 52 publications

Conformational Studies of RS-66271, an Analog of Parathyroid Hormone-Related Protein with Pronounced Bone Anabolic Activity

Conformational Studies of RS-66271, an Analog of Parathyroid Hormone-Related Protein with Pronounced Bone Anabolic Activity

Conformational studies of parathyroid hormone (PTH)/PTH-related protein (PTHrP) point-mutated hybrids

Conformational studies of parathyroid hormone (PTH)/PTH-related protein (PTHrp) chimeric peptides

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