Short Communication
NMR assignments of tryptophan residue in apo and holo LBD-rVDR
Wanda Sicinska,
William M. Westler,
Hector F. DeLuca,
Wanda Sicinska
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin
Search for more papers by this authorWilliam M. Westler
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin
Search for more papers by this authorCorresponding Author
Hector F. DeLuca
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin
Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706===Search for more papers by this authorWanda Sicinska,
William M. Westler,
Hector F. DeLuca,
Wanda Sicinska
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin
Search for more papers by this authorWilliam M. Westler
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin
Search for more papers by this authorCorresponding Author
Hector F. DeLuca
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin
Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706===Search for more papers by this authorAbstract
Binding sites in the full-length, ligand-binding domain of rat vitamin D receptor (LBD-rVDR) for an active hormone derived from vitamin D (1α,25-dihydroxyvitamin D3) and three of its C-2 substituted analogs were compared by nuclear magnetic resonance (NMR) spectroscopy. Specific residue labeled with [UL]-15N2 Trp allowed assignment of the side-chain Hϵ1 and Nε1 resonances of the single tryptophan residue at position 282 in LBD-rVDR. Comparison of 1H[15N] Heteronuclear Single Quantum Correlation (HSQC) spectra of apo and holo LBD-rVDR revealed that the position of the Trp282 Hε1 and Nε1 signals are sensitive to the presence of the ligand in the receptor cavity. Binding of the ligands to LBD-rVDR results in a shift of both Trp Hε1 and Nε1 resonances to lower frequencies. The results indicate that the interaction between the ligands and Trp282 is not responsible for differences in calcemic activity observed in vitamin D analogs. Proteins 2005. © 2005 Wiley-Liss, Inc.
REFERENCES
- 1 Bouillon R, Okamura WH, Norman AW. Structure-function relationships in the vitamin D endocrine system. Endocr Rev 1995; 16: 200–257.
- 2 DeLuca HF, Zierold C. Mechanism and functions of vitamin D. Nutr Rev 1998; 56: S4–S10.
- 3 Yamada S, Shimizu M, Yamamoto K. Structure-function relationships of vitamin D including ligand recognition by the vitamin D receptor. Med Res Rev 2003; 23: 89–115.
- 4 Griffin MD, Xing N, Kumar R. Vitamin D and its analogs as regulators of immune activation and antigen presentation. Annu Rev Nutr 2003; 23: 117–145.
- 5 Vaisanen S, Perakyla M, Karkkainen JI, Steinmeyer A, Carlberg C. Critical role of helix 12 of the vitamin D3 receptor for the partial agonism of carboxylic ester antagonists. J Mol Biol 2002; 315: 229–238.
- 6 Issa LL, Leong GM, Sutherland RL, Eisman JA. Vitamin D analogue-specific recruitment of vitamin D receptor coactivators. J Bone Miner Res 2002; 17: 879–890.
- 7 Choi M, Yamamoto K, Itoh T, Makishima M, Mangelsdorf DJ, Moras D, DeLuca HF, Yamada S. Interaction between vitamin D receptor and vitamin D ligands: two-dimensional alanine scanning mutational analysis. Chem and Biol 2003; 10: 261–270.
- 8 Yamamoto H, Shevde NK, Warrier A, Plum LA, DeLuca HF, Pike JW. 2-Methylene-19-nor-(20S)-1,25-dihydroxyvitamin D3 potently stimulates gene-specific DNA binding of the vitamin D receptor in osteoblasts. J Biol Chem 2003; 278: 31756–31765.
- 9 Herdick M, Carlberg C. Agonist-triggered modulation of the activated and silent state of the vitamin D3 receptor by interaction with co-repressors and co-activators. J Mol Biol 2000; 304: 793–801.
- 10 Yamamoto K, Ooizumi H, Umesono K, Verstuyf A, Bouillon R, DeLuca HF, Shinki T, Suda T, Yamada S. Three-dimensional structure-function relationship of vitamin D: side chain location and various activities. Bioorg Med Chem Lett 1999; 9: 1041–1046.
- 11 DeLuca HF, Sicinski RR, Gowlugari S, Plum LA, Clagett-Dame M. 1α-hydroxy-2-methylene-19-nor-homopregnacalciferol and its uses. US Patent 2002;6,440,953.
- 12 Sicinski RR, Prahl JM, Smith CM, DeLuca HF. New 1α,25-dihydroxy-19-norvitamin D3 compounds of high biological activity: synthesis and biological evaluation of 2-hydroxymethyl, 2-methyl, and 2-methylene analogues. J Med Chem 1998; 41: 4662–4674.
- 13 Sicinski RR, Rotkiewicz P, Kolinski A, Sicinska W, Prahl JM, Smith CM, DeLuca HF. 2-Ethyl and 2-ethylidene analogues of 1α,25-dihydroxy-19-norvitamin D3: synthesis, conformational analysis, biological activities, and docking to the modeled rVDR ligand binding domain. J Med Chem 2002; 45: 3366–3380.
- 14 Shevde NK, Plum LA, Clagett-Dame M, Yamamoto H, Pike JW, DeLuca HF. A potent analog of 1α,25-dihydroxyvitamin D3 selectively induces bone formation. Proc Natl Acad Sci USA 2002; 99: 13487–13491.
- 15 Suhara Y, Nihei K, Tanigawa H, Fujishima T, Konno K, Nakagawa K, Okano T, Takayama H. Syntheses and biological evaluation of novel 2α-substituted 1α,25-dihydroxyvitamin D3 analogues. Bioorg Med Chem Lett 2000; 10: 1129–1132.
- 16 Rochel N, Wurtz JM, Mitschler A, Klaholz B, Moras D. The crystal structure of the nuclear receptor for vitamin D bound to its natural ligand. Mol Cell 2000; 5: 173–179.
- 17 Tocchini-Valentini G, Rochel N, Wurtz JM, Mitschler A, Moras D. Crystal structures of the vitamin D receptor complexed to superagonist 20-epi ligands. Proc Natl Acad Sci USA 2001; 98: 5491–5496.
- 18 Vanhooke JL, Benning MM, Bauer CB, Pike JW, DeLuca HF Molecular structure of the rat vitamin D receptor ligand binding domain complexed with 2-carbon-substituted vitamin D3 hormone analogs and an LXXLL-containing coactivator peptide. Biochem 2004; 43: 4101–4110.
- 19 Okamura WH, Do S, Kim H, Jeganathan S, Vu T, Zhu GD, Norman AW. Conformationally restricted mimics of vitamin D rotamers. Steroids 2001; 66: 239–247.
- 20 Suwinska K, Kutner A. Crystal and molecular structure of 1,25-dihydroxycholecalciferol. Acta Cryst 1996; B52: 550–554.
- 21 Vaisanen S, Ryhanen S, Saarela JTA, Perakyla M, Andersin T, Maenpaa PH. Structurally and functionally important amino acids of the agonistic conformation of the human vitamin D receptor. Mol Pharm 2002; 62: 788–794.
- 22 Nguyen TM, Adiceam P, Kottler ML, Guillozo H, Rizk-Rabin M, Brouillard F, Lagier P, Palix C, Garnier JM, Garabedian M. Tryptophan missense mutation in the ligand-binding domain of the vitamin D receptor causes severe resistance to 1,25-dihydroxy vitamin D. J Bone Miner Res 2002; 17: 1728–1737.
- 23 Ray R, Swamy N, MacDonald PN, Ray S, Haussler MR, Holick MF. Affinity labeling of the 1α,25-dihydroxyvitamin D3 receptor. J Biol Chem 1996; 271, 2012–2017.
- 24 Strugnell SA, Hill JJ, McCaslin DR, Wiefling BA, Royer CA, DeLuca HF. Bacterial expression and characterization of the ligand binding domain of the vitamin D receptor. Arch Biochem Biophys 1999; 364: 42–52.
- 25 Nayeri S, Carsten C. Functional conformations of the nuclear 1α,25-dihydroxyvitamin D3 receptor. Biochem J 1997; 327: 561–568.
- 26 Strugnell SA, DeLuca HF. The vitamin D receptor-structure and transcriptional activation. Proc Soc Exp Biol Med 1997; 215: 223–228.
- 27 Solomon C, Macoritto M, Gao XL, White JH, Kremer R. The unique tryptophan residue of the vitamin D receptor is critical for ligand binding and transcriptional activation. J Bone Miner Res 2001; 16: 39–45.
- 28 Sicinska W, Rotkiewicz P, DeLuca HF. Model of three-dimensional structure of VDR bound with vitamin D3 analogs substituted at carbon 2. Proceedings of the 12th Workshop on Vitamin D in J Steroid Biochem Mol Biol 2004; 89–90: 107–110.
- 29 Mohammadi F, Prentice GA, Merril AR. Protein-protein interaction using tryptophan analogues: novel spectroscopic probes for toxin-elongation factor-2 interactions. Biochem 2001; 40: 10273–10283.
- 30 Cheng H, Westler WM, Xia B, Oh BH, Markley JL. Protein expression, selective isotope labeling, and analysis of hyperfine-shifted NMR signals of anabaena 7120 vegetative [2Fe-2S] ferrodoxin. Arch Biochem Biophys 1995; 316: 619–634.
- 31 Dame MC, Pierce EA, Prahl JM, Hayes CE, DeLuca HF. Monoclonal antibodies to the porcine intestinal receptor foe 1,25-dihydroxyvitamin D3: Interaction with distinct receptor domains. Biochem 1986; 25: 4523–4534.
- 32 Gallagher SR, One-dimensional electrophoresis using nondenaturating conditions. In: GP Taylor, editor. Current protocols in protein science. Somerset, NY: Wiley, 1995–2000. p 10.3.
- 33 Mori S, Abeygunawardana C, Johnson MO, Vanzijl PCM. Improved sensitivity of HSQC spectra of exchanging protons at short interscan delays using a new fast HSQC (FHSQC) detection scheme that avoids water saturation. J Magn Reson Series B 1995; 108: 94–8.
- 34 Markley J, Bax A, Arata Y, Hilbers CW, Kaptein R, Sykes BD, Wright PE, Wuthrich K. Recommendations for the presentation of NMR structures of proteins and nucleic acids. Pure Appl Chem 1998; 70: 117–142.
- 35 Wider G, Wuthrich K. NMR spectroscopy of large molecules and multinuclear assemblies in solution. Curr Opin Struct Biol 1999; 9: 594–601.
- 36 Markley JL, Kainosho M. Stable isotope labelling and resonance assignments in larger proteins. In: GCK Roberts, editor. NMR of macromolecules. A practical approach. Oxford: Oxford University Press, 1993. p 101–152.
- 37 Gunther H. NMR Spectroscopy. New York: Wiley; 1980. 75 p.
- 38 Melander W, Horvath C. Salt effects on hydrophobic interactions in precipitation and chromatography of proteins: an interpretation of the lyotropic series. Arch Bioch Biophys 1977; 183: 200–215.
- 39 Symons MCR. Water structure and reactivity. Acc Chem Res 1981; 14: 179–187.
- 40 Witanowski M, Sicinska W, Grabowski Z, Webb GA. Pyrrole and N-methylpyrrole as models for solvent polarity and solute-to-solvent hydrogen bonding effects on nitrogen NMR shielding. J Magn Reson 1993: 104A: 310–314.
- 41 Witanowski M, Stefaniak L, Webb GA. Nitrogen NMR spectroscopy. In: GA Webb, editor. Annual reports on NMR spectroscopy, vol 25. San Diego: Academic Press; 1993. p 225.
- 42 Witanowski M, Stefaniak L, Webb GA. Nitrogen NMR spectroscopy. In: GA Webb, editor. Annual reports on NMR spectroscopy, vol 25. San Diego: Academic Press; 1993. p 50.
- 43 Lee W, Noy N. Interactions of RXR with coactivators are differentially mediated by helix 11 of receptor's ligand binding domain. Biochem 2001; 41: 2500–2508.
- 44 Barry JB, Leong GM, Church WB, Issa LL, Eisman JA, Gardiner EM. Interactions of SKIP/NCoA-62, TFIIB, and Retinoid X receptor with vitamin D receptor helix H10 residues. J Biol Chem 2003; 278: 8224–8228.
- 45 Pathrose P, Barmina O, Chang CY, McDonnell DP, Shevde NK, Pike JW. Inhibition of 1,25-dihydroxyvitamin D3-dependent transcription by synthetic LXXLL peptide antagonists that target the activation domains of the vitamin D and retinoid X receptors. J Bone Miner Res 2002; 17: 2196–2205.
- 46 He B, Bowen NT, Minges JT, Wilson EM. Androgen-induced NH2- and COOH-terminal interaction inhibits p160 coactivator recruitment by activation function 2. J Biol Chem 2001; 276: 42293–42301.
- 47 Gonzales-Avion XC, Mourino A. Functionalization at C-12 of 1α,25-dihydroxyvitamin D3 strongly modulates the affinity for the vitamin D receptor (VDR). Org Lett 2003; 5: 2291–2293.
- 48 Sicinski RR, Perlman KL, Prahl J, Smith C, DeLuca HF. Synthesis and biological activity of 1α,25-dihydroxy-18-norvitamin D3 and 1α,25-dihydroxy-18,19-dinorvitamin D3. J Med Chem 1995; 39: 4497–4506.
- 49 Sicinski RR, Prahl JM, Smith CM, DeLuca HF. New highly calcemic 1α,25-dihydroxy-19-norvitmain D3 compounds with modified side chain: 26,27-dihomo- and 26,27-dimethylene analogs in 20S-series. Steroids 2002; 67: 247–256.
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