메뉴 건너뛰기




Volumn 411, Issue 4, 2011, Pages 896-909

Parvulin 17 promotes microtubule assembly by its peptidyl-prolyl cis/trans isomerase activity

Author keywords

microtubules; parvulin; peptide microarray; PPIase; tubulin

Indexed keywords

BETA TUBULIN; GUANOSINE TRIPHOSPHATE; MICROTUBULE ASSOCIATED PROTEIN; PARVULIN 17; PEPTIDYLPROLYL ISOMERASE; TUBULIN; UNCLASSIFIED DRUG;

EID: 80051671474     PISSN: 00222836     EISSN: 10898638     Source Type: Journal    
DOI: 10.1016/j.jmb.2011.06.040     Document Type: Article
Times cited : (18)

References (58)
  • 1
    • 0028124244 scopus 로고
    • Confirmation of the existence of a third family among peptidyl-prolyl cis/trans isomerases amino acid sequence and recombinant production of parvulin
    • DOI 10.1016/0014-5793(94)00932-5
    • Rahfeld J.U., Rucknagel K.P., Schelbert B., Ludwig B., Hacker J., Mann K., and Fischer G. Confirmation of the existence of a third family among peptidyl-prolyl cis/trans isomerases. Amino acid sequence and recombinant production of parvulin FEBS Lett. 352 1994 180 184 (Pubitemid 24298414)
    • (1994) FEBS Letters , vol.352 , Issue.2 , pp. 180-184
    • Fischer, G.1
  • 2
    • 28444439801 scopus 로고    scopus 로고
    • Insights into the catalytic mechanism of peptidyl prolyl cis/trans isomerases
    • Fanghanel J., and Fischer G. Insights into the catalytic mechanism of peptidyl prolyl cis/trans isomerases Front. Biosci. 9 2004 3453 3478
    • (2004) Front. Biosci. , vol.9 , pp. 3453-3478
    • Fanghanel, J.1    Fischer, G.2
  • 3
    • 57149093203 scopus 로고    scopus 로고
    • FKBP family proteins: Immunophilins with versatile biological functions
    • Kang C.B., Hong Y., Dhe-Paganon S., and Yoon H.S. FKBP family proteins: immunophilins with versatile biological functions Neurosignals 16 2008 318 325
    • (2008) Neurosignals , vol.16 , pp. 318-325
    • Kang, C.B.1    Hong, Y.2    Dhe-Paganon, S.3    Yoon, H.S.4
  • 4
    • 0029852803 scopus 로고    scopus 로고
    • Chaperone function of Hsp90-associated proteins
    • DOI 10.1126/science.274.5293.1715
    • Bose S., Weikl T., Bugl H., and Buchner J. Chaperone function of Hsp90-associated proteins Science 274 1996 1715 1717 (Pubitemid 26414904)
    • (1996) Science , vol.274 , Issue.5293 , pp. 1715-1717
    • Bose, S.1    Weikl, T.2    Bugl, H.3    Buchner, J.4
  • 5
    • 0040017910 scopus 로고    scopus 로고
    • Function of WW domains as phosphoserine- or phosphothreonine-binding modules
    • Lu P.J., Zhou X.Z., Shen M., and Lu K.P. Function of WW domains as phosphoserine- or phosphothreonine-binding modules Science 283 1999 1325 1328
    • (1999) Science , vol.283 , pp. 1325-1328
    • Lu, P.J.1    Zhou, X.Z.2    Shen, M.3    Lu, K.P.4
  • 7
    • 4344679004 scopus 로고    scopus 로고
    • Solution structure of Escherichia coli Par10: The prototypic member of the Parvulin family of peptidyl-prolyl cis/trans isomerases
    • DOI 10.1110/ps.04756704
    • Kuhlewein A., Voll G., Hernandez Alvarez B., Kessler H., Fischer G., Rahfeld J.U., and Gemmecker G. Solution structure of Escherichia coli Par10: the prototypic member of the parvulin family of peptidyl-prolyl cis/trans isomerases Protein Sci. 13 2004 2378 2387 (Pubitemid 39128858)
    • (2004) Protein Science , vol.13 , Issue.9 , pp. 2378-2387
    • Kuhlewein, A.1    Voll, G.2    Alvarez, B.H.3    Kessler, H.4    Fischer, G.5    Rahfeld, J.-U.6    Gemmecker, G.7
  • 8
    • 0034714176 scopus 로고    scopus 로고
    • NMR solution structure of hPar14 reveals similarity to the peptidyl prolyl cis/trans isomerase domain of the mitotic regulator hPin1 but indicates a different functionality of the protein
    • Sekerina E., Rahfeld J.U., Muller J., Fanghanel J., Rascher C., Fischer G., and Bayer P. NMR solution structure of hPar14 reveals similarity to the peptidyl prolyl cis/trans isomerase domain of the mitotic regulator hPin1 but indicates a different functionality of the protein J. Mol. Biol. 301 2000 1003 1017
    • (2000) J. Mol. Biol. , vol.301 , pp. 1003-1017
    • Sekerina, E.1    Rahfeld, J.U.2    Muller, J.3    Fanghanel, J.4    Rascher, C.5    Fischer, G.6    Bayer, P.7
  • 9
    • 70350491197 scopus 로고    scopus 로고
    • A library of fluorescent peptides for exploring the substrate specificities of prolyl isomerases
    • Zoldak G., Aumuller T., Lucke C., Hritz J., Oostenbrink C., Fischer G., and Schmid F.X. A library of fluorescent peptides for exploring the substrate specificities of prolyl isomerases Biochemistry 48 2009 10423 10436
    • (2009) Biochemistry , vol.48 , pp. 10423-10436
    • Zoldak, G.1    Aumuller, T.2    Lucke, C.3    Hritz, J.4    Oostenbrink, C.5    Fischer, G.6    Schmid, F.X.7
  • 10
    • 0035947078 scopus 로고    scopus 로고
    • Pin1 acts catalytically to promote a conformational change in Cdc25
    • DOI 10.1016/S1097-2765(01)00245-3
    • Stukenberg P.T., and Kirschner M.W. Pin1 acts catalytically to promote a conformational change in Cdc25 Mol. Cell 7 2001 1071 1083 (Pubitemid 32525753)
    • (2001) Molecular Cell , vol.7 , Issue.5 , pp. 1071-1083
    • Stukenberg P.Todd1    Kirschner, M.W.2
  • 11
    • 0033638180 scopus 로고    scopus 로고
    • Pin1-dependent prolyl isomerization regulates dephosphorylation of Cdc25C and tau proteins
    • Zhou X.Z., Kops O., Werner A., Lu P.J., Shen M., and Stoller G. Pin1-dependent prolyl isomerization regulates dephosphorylation of Cdc25C and tau proteins Mol. Cell 6 2000 873 883
    • (2000) Mol. Cell , vol.6 , pp. 873-883
    • Zhou, X.Z.1    Kops, O.2    Werner, A.3    Lu, P.J.4    Shen, M.5    Stoller, G.6
  • 13
    • 0033600242 scopus 로고    scopus 로고
    • The prolyl isomerase Pin1 restores the function of Alzheimer-associated phosphorylated tau protein
    • DOI 10.1038/21650
    • Lu P.J., Wulf G., Zhou X.Z., Davies P., and Lu K.P. The prolyl isomerase Pin1 restores the function of Alzheimer-associated phosphorylated tau protein Nature 399 1999 784 788 (Pubitemid 29293174)
    • (1999) Nature , vol.399 , Issue.6738 , pp. 784-788
    • Lu, P.-J.1    Wulf, G.2    Zhou, X.Z.3    Davies, P.4    Lu, K.P.5
  • 14
    • 0036382919 scopus 로고    scopus 로고
    • The N-terminal basic domain of human parvulin hPar14 is responsible for the entry to the nucleus and high-affinity DNA-binding
    • Surmacz T.A., Bayer E., Rahfeld J.U., Fischer G., and Bayer P. The N-terminal basic domain of human parvulin hPar14 is responsible for the entry to the nucleus and high-affinity DNA-binding J. Mol. Biol. 321 2002 235 247
    • (2002) J. Mol. Biol. , vol.321 , pp. 235-247
    • Surmacz, T.A.1    Bayer, E.2    Rahfeld, J.U.3    Fischer, G.4    Bayer, P.5
  • 16
    • 71049184757 scopus 로고    scopus 로고
    • Parvulin (Par14), a peptidyl-prolyl cis-trans isomerase, is a novel rRNA processing factor that evolved in the metazoan lineage
    • Fujiyama-Nakamura S., Yoshikawa H., Homma K., Hayano T., Tsujimura-Takahashi T., and Izumikawa K. Parvulin (Par14), a peptidyl-prolyl cis-trans isomerase, is a novel rRNA processing factor that evolved in the metazoan lineage Mol. Cell. Proteomics 8 2009 1552 1565
    • (2009) Mol. Cell. Proteomics , vol.8 , pp. 1552-1565
    • Fujiyama-Nakamura, S.1    Yoshikawa, H.2    Homma, K.3    Hayano, T.4    Tsujimura-Takahashi, T.5    Izumikawa, K.6
  • 18
    • 0038690213 scopus 로고    scopus 로고
    • Phosphorylation of the N-terminal domain regulates subcellular localization and DNA binding properties of the peptidyl-prolyl cis/trans isomerase hPar14
    • DOI 10.1016/S0022-2836(03)00713-7
    • Reimer T., Weiwad M., Schierhorn A., Ruecknagel P.K., Rahfeld J.U., Bayer P., and Fischer G. Phosphorylation of the N-terminal domain regulates subcellular localization and DNA binding properties of the peptidyl-prolyl cis/trans isomerase hPar14 J. Mol. Biol. 330 2003 955 966 (Pubitemid 36818894)
    • (2003) Journal of Molecular Biology , vol.330 , Issue.5 , pp. 955-966
    • Reimer, T.1    Weiwad, M.2    Schierhorn, A.3    Ruecknagel, P.-K.4    Rahfeld, J.-U.5    Bayer, P.6    Fischer, G.7
  • 19
    • 35448936378 scopus 로고    scopus 로고
    • The DNA binding parvulin Par17 is targeted to the mitochondrial matrix by a recently evolved prepeptide uniquely present in Hominidae
    • Kessler D., Papatheodorou P., Stratmann T., Dian E.A., Hartmann-Fatu C., and Rassow J. The DNA binding parvulin Par17 is targeted to the mitochondrial matrix by a recently evolved prepeptide uniquely present in Hominidae BMC Biol. 5 2007 37
    • (2007) BMC Biol. , vol.5 , pp. 37
    • Kessler, D.1    Papatheodorou, P.2    Stratmann, T.3    Dian, E.A.4    Hartmann-Fatu, C.5    Rassow, J.6
  • 20
    • 33645349063 scopus 로고    scopus 로고
    • Characterization of novel elongated parvulin isoforms that are ubiquitously expressed in human tissues and originate from alternative transcription initiation
    • Mueller J.W., Kessler D., Neumann D., Stratmann T., Papatheodorou P., Hartmann-Fatu C., and Bayer P. Characterization of novel elongated parvulin isoforms that are ubiquitously expressed in human tissues and originate from alternative transcription initiation BMC Mol. Biol. 7 2006 9
    • (2006) BMC Mol. Biol. , vol.7 , pp. 9
    • Mueller, J.W.1    Kessler, D.2    Neumann, D.3    Stratmann, T.4    Papatheodorou, P.5    Hartmann-Fatu, C.6    Bayer, P.7
  • 21
    • 0034743675 scopus 로고    scopus 로고
    • Structural insight into microtubule function
    • Nogales E. Structural insight into microtubule function Annu. Rev. Biophys. Biomol. Struct. 30 2001 397 420
    • (2001) Annu. Rev. Biophys. Biomol. Struct. , vol.30 , pp. 397-420
    • Nogales, E.1
  • 22
    • 0021982448 scopus 로고
    • Actin and tubulin polymerization: The use of kinetic methods to determine mechanism
    • Frieden C. Actin and tubulin polymerization: the use of kinetic methods to determine mechanism Annu. Rev. Biophys. Biophys. Chem. 14 1985 189 210
    • (1985) Annu. Rev. Biophys. Biophys. Chem. , vol.14 , pp. 189-210
    • Frieden, C.1
  • 25
    • 0019315916 scopus 로고
    • Evidence for two growth steps in microtubule polymerization
    • Barton J.S., and Riazi G.H. Evidence for two growth steps in microtubule polymerization Biochim. Biophys. Acta 630 1980 392 401
    • (1980) Biochim. Biophys. Acta , vol.630 , pp. 392-401
    • Barton, J.S.1    Riazi, G.H.2
  • 26
    • 0030783012 scopus 로고    scopus 로고
    • 2S complex with two tubulin molecules
    • DOI 10.1021/bi971491b
    • Jourdain L., Curmi P., Sobel A., Pantaloni D., and Carlier M.F. A tubulin-sequestering protein which forms a ternary T2S complex with two tubulin molecules Biochemistry 36 1997 10817 10821 (Pubitemid 27396235)
    • (1997) Biochemistry , vol.36 , Issue.36 , pp. 10817-10821
    • Jourdain, L.1    Curmi, P.2    Sobel, A.3    Pantaloni, D.4    Carlier, M.-F.5
  • 27
    • 0024556885 scopus 로고
    • A common amino acid sequence in 190-kDa microtubule-associated protein and tau for the promotion of microtubule assembly
    • Aizawa H., Kawasaki H., Murofushi H., Kotani S., Suzuki K., and Sakai H. A common amino acid sequence in 190-kDa microtubule-associated protein and tau for the promotion of microtubule assembly J. Biol. Chem. 264 1989 5885 5890 (Pubitemid 19098057)
    • (1989) Journal of Biological Chemistry , vol.264 , Issue.10 , pp. 5885-5890
    • Aizawa, H.1    Kawasaki, H.2    Murofushi, H.3    Kotani, S.4    Suzuki, K.5    Sakai, H.6
  • 28
    • 1642499478 scopus 로고    scopus 로고
    • Cypin regulates dendrite patterning in hippocampal neurons by promoting microtubule assembly
    • DOI 10.1038/nn1179
    • Akum B.F., Chen M., Gunderson S.I., Riefler G.M., Scerri-Hansen M.M., and Firestein B.L. Cypin regulates dendrite patterning in hippocampal neurons by promoting microtubule assembly Nat. Neurosci. 7 2004 145 152 (Pubitemid 38129790)
    • (2004) Nature Neuroscience , vol.7 , Issue.2 , pp. 145-152
    • Akum, B.F.1    Chen, M.2    Gunderson, S.I.3    Riefler, G.M.4    Scerri-Hansen, M.M.5    Firestein, B.L.6
  • 29
    • 0030048731 scopus 로고    scopus 로고
    • Identification of a protein that interacts with tubulin dimers and increases the catastrophe rate of microtubules
    • DOI 10.1016/S0092-8674(00)81037-5
    • Belmont L.D., and Mitchison T.J. Identification of a protein that interacts with tubulin dimers and increases the catastrophe rate of microtubules Cell 84 1996 623 631 (Pubitemid 26071742)
    • (1996) Cell , vol.84 , Issue.4 , pp. 623-631
    • Belmont, L.D.1    Mitchison, T.J.2
  • 31
    • 0032981664 scopus 로고    scopus 로고
    • Identification of the binding region of basic calponin on α and β tubulins
    • Fujii T., and Koizumi Y. Identification of the binding region of basic calponin on alpha and beta tubulins J. Biochem. 125 1999 869 875 (Pubitemid 29253554)
    • (1999) Journal of Biochemistry , vol.125 , Issue.5 , pp. 869-875
    • Fujii, T.1    Koizumi, Y.2
  • 33
    • 0030878593 scopus 로고    scopus 로고
    • The transport and binding of taxol
    • DOI 10.1016/S0306-3623(97)89716-9
    • Parekh H., and Simpkins H. The transport and binding of taxol Gen. Pharmacol. 29 1997 167 172 (Pubitemid 27312365)
    • (1997) General Pharmacology , vol.29 , Issue.2 , pp. 167-172
    • Parekh, H.1    Simpkins, H.2
  • 34
    • 0026087554 scopus 로고
    • Effects of 10 known or suspected spindle poisons in the in vitro porcine brain tubulin assembly assay
    • Brunner M., Albertini S., and Wurgler F.E. Effects of 10 known or suspected spindle poisons in the in vitro porcine brain tubulin assembly assay Mutagenesis 6 1991 65 70
    • (1991) Mutagenesis , vol.6 , pp. 65-70
    • Brunner, M.1    Albertini, S.2    Wurgler, F.E.3
  • 36
    • 0016648839 scopus 로고
    • Fluorometric assay of tubulin-colchicine complex
    • Arai T., and Okuyama T. Fluorometric assay of tubulin-colchicine complex Anal. Biochem. 69 1975 443 450
    • (1975) Anal. Biochem. , vol.69 , pp. 443-450
    • Arai, T.1    Okuyama, T.2
  • 37
    • 0032005075 scopus 로고    scopus 로고
    • Tubulin and microtubule structure
    • DOI 10.1016/S0955-0674(98)80082-3
    • Downing K.H., and Nogales E. Tubulin and microtubule structure Curr. Opin. Cell Biol. 10 1998 16 22 (Pubitemid 28066117)
    • (1998) Current Opinion in Cell Biology , vol.10 , Issue.1 , pp. 16-22
    • Downing, K.H.1    Nogales, E.2
  • 39
    • 34248225770 scopus 로고    scopus 로고
    • Structure and thermodynamics of the tubulin-stathmin interaction
    • DOI 10.1016/j.jsb.2006.07.018, PII S1047847706002437, Structural Analysis of Supramolecular Assembles by Hybrid Methods
    • Steinmetz M.O. Structure and thermodynamics of the tubulin-stathmin interaction J. Struct. Biol. 158 2007 137 147 (Pubitemid 46719987)
    • (2007) Journal of Structural Biology , vol.158 , Issue.2 , pp. 137-147
    • Steinmetz, M.O.1
  • 41
    • 34548485298 scopus 로고    scopus 로고
    • The immunophilin FKBP52 specifically binds to tubulin and prevents microtubule formation
    • DOI 10.1096/fj.06-7667com
    • Chambraud B., Belabes H., Fontaine-Lenoir V., Fellous A., and Baulieu E.E. The immunophilin FKBP52 specifically binds to tubulin and prevents microtubule formation FASEB J. 21 2007 2787 2797 (Pubitemid 47372746)
    • (2007) FASEB Journal , vol.21 , Issue.11 , pp. 2787-2797
    • Chambraud, B.1    Belabes, H.2    Fontaine-Lenoir, V.3    Fellous, A.4    Baulieu, E.E.5
  • 43
    • 44049092273 scopus 로고    scopus 로고
    • Conformational analysis of the carboxy-terminal tails of human beta-tubulin isotypes
    • Luchko T., Huzil J.T., Stepanova M., and Tuszynski J. Conformational analysis of the carboxy-terminal tails of human beta-tubulin isotypes Biophys. J. 94 2008 1971 1982
    • (2008) Biophys. J. , vol.94 , pp. 1971-1982
    • Luchko, T.1    Huzil, J.T.2    Stepanova, M.3    Tuszynski, J.4
  • 44
    • 33947713961 scopus 로고    scopus 로고
    • Recognition of C-terminal amino acids in tubulin by pore loops in Spastin is important for microtubule severing
    • DOI 10.1083/jcb.200610072
    • White S.R., Evans K.J., Lary J., Cole J.L., and Lauring B. Recognition of C-terminal amino acids in tubulin by pore loops in spastin is important for microtubule severing J. Cell Biol. 176 2007 995 1005 (Pubitemid 46506972)
    • (2007) Journal of Cell Biology , vol.176 , Issue.7 , pp. 995-1005
    • White, S.R.1    Evans, K.J.2    Lary, J.3    Cole, J.L.4    Lauring, B.5
  • 48
    • 33645680245 scopus 로고    scopus 로고
    • Nanomolar inhibitors of the peptidyl prolyl cis/trans isomerase Pin1 from combinatorial peptide libraries
    • Wildemann D., Erdmann F., Hernandez Alvarez B., Stoller G., Zhou X.Z., and Fanghänel J. Nanomolar inhibitors of the peptidyl prolyl cis/trans isomerase Pin1 from combinatorial peptide libraries J. Med. Chem. 49 2006 2147 2150
    • (2006) J. Med. Chem. , vol.49 , pp. 2147-2150
    • Wildemann, D.1    Erdmann, F.2    Hernandez Alvarez, B.3    Stoller, G.4    Zhou, X.Z.5    Fanghänel, J.6
  • 49
    • 33748487414 scopus 로고    scopus 로고
    • Myelin basic protein: A multifunctional protein
    • DOI 10.1007/s00018-006-6094-7
    • Boggs J.M. Myelin basic protein: a multifunctional protein Cell. Mol. Life Sci. 63 2006 1945 1961 (Pubitemid 44359737)
    • (2006) Cellular and Molecular Life Sciences , vol.63 , Issue.17 , pp. 1945-1961
    • Boggs, J.M.1
  • 50
    • 1642401199 scopus 로고    scopus 로고
    • Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain
    • DOI 10.1038/nature02393
    • Ravelli R.B., Gigant B., Curmi P.A., Jourdain I., Lachkar S., Sobel A., and Knossow M. Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain Nature 428 2004 198 202 (Pubitemid 38374318)
    • (2004) Nature , vol.428 , Issue.6979 , pp. 198-202
    • Ravelli, R.B.G.1    Gigant, B.2    Curmi, P.A.3    Jourdain, I.4    Lachkar, S.5    Sobel, A.6    Knossow, M.7
  • 51
    • 12344320203 scopus 로고    scopus 로고
    • Microtubule plus-end-tracking proteins: Mechanisms and functions
    • DOI 10.1016/j.ceb.2004.11.001, PII S095506740400167X
    • Akhmanova A., and Hoogenraad C.C. Microtubule plus-end-tracking proteins: mechanisms and functions Curr. Opin. Cell Biol. 17 2005 47 54 (Pubitemid 40138087)
    • (2005) Current Opinion in Cell Biology , vol.17 , Issue.1 , pp. 47-54
    • Akhmanova, A.1    Hoogenraad, C.C.2
  • 52
    • 0028021695 scopus 로고
    • Tubulin GTP hydrolysis influences the structure, mechanical properties, and kinesin-driven transport of microtubules
    • Vale R.D., Coppin C.M., Malik F., Kull F.J., and Milligan R.A. Tubulin GTP hydrolysis influences the structure, mechanical properties, and kinesin-driven transport of microtubules J. Biol. Chem. 269 1994 23769 23775 (Pubitemid 24293587)
    • (1994) Journal of Biological Chemistry , vol.269 , Issue.38 , pp. 23769-23775
    • Vale, R.D.1    Coppin, C.M.2    Malik, F.3    Kull, F.J.4    Milligan, R.A.5
  • 54
    • 0025202947 scopus 로고
    • Associations between beta-tubulin and mitochondria in adult isolated heart myocytes as shown by immunofluorescence and immunoelectron microscopy
    • Saetersdal T., Greve G., and Dalen H. Associations between beta-tubulin and mitochondria in adult isolated heart myocytes as shown by immunofluorescence and immunoelectron microscopy Histochemistry 95 1990 1 10
    • (1990) Histochemistry , vol.95 , pp. 1-10
    • Saetersdal, T.1    Greve, G.2    Dalen, H.3
  • 56
    • 0033039865 scopus 로고    scopus 로고
    • Identification and characterization of a 14 kDa human protein as a novel parvulin-like peptidyl prolyl cis/trans isomerase
    • DOI 10.1016/S0014-5793(99)00239-2, PII S0014579399002392
    • Uchida T., Fujimori F., Tradler T., Fischer G., and Rahfeld J.U. Identification and characterization of a 14 kDa human protein as a novel parvulin-like peptidyl prolyl cis/trans isomerase FEBS Lett. 446 1999 278 282 (Pubitemid 29132743)
    • (1999) FEBS Letters , vol.446 , Issue.2-3 , pp. 278-282
    • Uchida, T.1    Fujimori, F.2    Tradler, T.3    Fischer, G.4    Rahfeld, J.-U.5
  • 57
    • 0021710353 scopus 로고
    • Interaction of tubulin with chromatin proteins. H1 and core histones
    • Mithieux G., Alquier C., Roux B., and Rousset B. Interaction of tubulin with chromatin proteins. H1 and core histones J. Biol. Chem. 259 1984 15523 15531 (Pubitemid 15190768)
    • (1984) Journal of Biological Chemistry , vol.259 , Issue.24 , pp. 15523-15531
    • Mithieux, G.1    Alquier, C.2    Roux, B.3    Rousset, B.4


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.