메뉴 건너뛰기




Volumn 19, Issue 3, 1996, Pages 455-466

Characterization of pectin methylesterase B, an outer membrane lipoprotein of Erwinia chrysanthemi 3937

Author keywords

[No Author keywords available]

Indexed keywords

ESTERASE; OUTER MEMBRANE PROTEIN; PECTINESTERASE;

EID: 0030039270     PISSN: 0950382X     EISSN: None     Source Type: Journal    
DOI: 10.1046/j.1365-2958.1996.389922.x     Document Type: Article
Times cited : (50)

References (52)
  • 1
    • 0026148980 scopus 로고
    • A gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of Brassica napus. Sequences in its 5′ flanking region are conserved in other pollen-specific promoters
    • Albani, D., Altosaar, I., Arnison, P.G., and Fabijanski, S.F. (1991) A gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of Brassica napus. Sequences in its 5′ flanking region are conserved in other pollen-specific promoters. Plant Mol Biol 16: 501-513.
    • (1991) Plant Mol Biol , vol.16 , pp. 501-513
    • Albani, D.1    Altosaar, I.2    Arnison, P.G.3    Fabijanski, S.F.4
  • 2
    • 0028156164 scopus 로고
    • Extracellular enzymes and pathogenesis of soft-rot Erwinia
    • Barras, F., van Gijsegem, F., and Chatterjee, A. (1994) Extracellular enzymes and pathogenesis of soft-rot Erwinia. Annu Rev Phytopathol 32: 201-234.
    • (1994) Annu Rev Phytopathol , vol.32 , pp. 201-234
    • Barras, F.1    Van Gijsegem, F.2    Chatterjee, A.3
  • 3
    • 0000424838 scopus 로고
    • Pathogenic behaviour of pectinase-defective Erwinia chrysanthemi mutants on different plants
    • Beaulieu, C., Boccara, M., and van Gijsegem, F. (1993) Pathogenic behaviour of pectinase-defective Erwinia chrysanthemi mutants on different plants. Mol Plant-Microb Interac 6: 197-202.
    • (1993) Mol Plant-Microb Interac , vol.6 , pp. 197-202
    • Beaulieu, C.1    Boccara, M.2    Van Gijsegem, F.3
  • 4
    • 0024707559 scopus 로고
    • Regulation and role in pathogenicity of Erwinia chrysanthemi 3937 pectin methylesterase
    • Boccara, M., and Chatain, V. (1988) Regulation and role in pathogenicity of Erwinia chrysanthemi 3937 pectin methylesterase. J Bacteriol 171: 4085-4087.
    • (1988) J Bacteriol , vol.171 , pp. 4085-4087
    • Boccara, M.1    Chatain, V.2
  • 5
    • 0026530575 scopus 로고
    • Platelet membrane phosphatidylinositol kinase activity. Triton X-100 effect provide evidence for intramicellar reaction
    • Boué, D., and Viratelle, O.M. (1992) Platelet membrane phosphatidylinositol kinase activity. Triton X-100 effect provide evidence for intramicellar reaction. Biochim Biophys Acta 1103: 120-126.
    • (1992) Biochim Biophys Acta , vol.1103 , pp. 120-126
    • Boué, D.1    Viratelle, O.M.2
  • 6
    • 0025605026 scopus 로고
    • Molecular cloning of the structural gene for exopolygalacturonate lyase from Erwinia chrysanthemi EC16 and characterization of the enzyme product
    • Brooks, A.D., He, S.Y., Gold, S., Keen, N.T., and Collmer, A. (1990) Molecular cloning of the structural gene for exopolygalacturonate lyase from Erwinia chrysanthemi EC16 and characterization of the enzyme product. J Bacteriol 172: 6950-6958.
    • (1990) J Bacteriol , vol.172 , pp. 6950-6958
    • Brooks, A.D.1    He, S.Y.2    Gold, S.3    Keen, N.T.4    Collmer, A.5
  • 7
    • 0024076255 scopus 로고
    • Collagen-like sequences stabilize homotrimers of a bacterial hydrolase
    • Charalamboulos, B.M., Keen, J.N., and McPherson, M.J. (1988) Collagen-like sequences stabilize homotrimers of a bacterial hydrolase. EMBO J 7: 2903-2909.
    • (1988) EMBO J , vol.7 , pp. 2903-2909
    • Charalamboulos, B.M.1    Keen, J.N.2    McPherson, M.J.3
  • 8
    • 0000980748 scopus 로고
    • The role of pectic enzymes in plant pathogenesis
    • Collmer, A., and Keen, N.T. (1986). The role of pectic enzymes in plant pathogenesis. Annu Rev Phytopathol 24: 383-409.
    • (1986) Annu Rev Phytopathol , vol.24 , pp. 383-409
    • Collmer, A.1    Keen, N.T.2
  • 9
    • 0020410210 scopus 로고
    • Renaturation and identification of periplasmic proteins in two-dimensional gels of Escherichia coli
    • Copeland, B.R., Richter, R.J., and Furlong, C.E. (1982) Renaturation and identification of periplasmic proteins in two-dimensional gels of Escherichia coli. J Biol Chem 257: 15065-15071.
    • (1982) J Biol Chem , vol.257 , pp. 15065-15071
    • Copeland, B.R.1    Richter, R.J.2    Furlong, C.E.3
  • 10
    • 0019063955 scopus 로고
    • Detection of pectic enzymes in pectin-acrylamide gels
    • Cruickshank, R.H., and Wade, G.C. (1980) Detection of pectic enzymes in pectin-acrylamide gels. Anal Biochem 107: 177-181.
    • (1980) Anal Biochem , vol.107 , pp. 177-181
    • Cruickshank, R.H.1    Wade, G.C.2
  • 11
    • 0015854977 scopus 로고
    • Solubilisation of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate
    • Filip, C., Fletcher, G., Wulf, J.L., and Earhart, C.F. (1973) Solubilisation of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol 115: 717-722.
    • (1973) J Bacteriol , vol.115 , pp. 717-722
    • Filip, C.1    Fletcher, G.2    Wulf, J.L.3    Earhart, C.F.4
  • 12
    • 0024744316 scopus 로고
    • Relationship between the pel genes of the pelADE cluster in Erwinia chrysanthemi B374
    • van Gijsegem, F. (1989) Relationship between the pel genes of the pelADE cluster in Erwinia chrysanthemi B374. Mol Microbiol 3: 1415-1424.
    • (1989) Mol Microbiol , vol.3 , pp. 1415-1424
    • Van Gijsegem, F.1
  • 13
    • 0020620358 scopus 로고
    • In vivo cloning of Erwinia chrysanthemi genes involved in the catabolism of hexuronates
    • van Gijsegem, F., and Toussaint, A. (1983) In vivo cloning of Erwinia chrysanthemi genes involved in the catabolism of hexuronates. J Bacteriol 154: 1227-1235.
    • (1983) J Bacteriol , vol.154 , pp. 1227-1235
    • Van Gijsegem, F.1    Toussaint, A.2
  • 14
    • 0025964225 scopus 로고
    • Molecular cloning, partial purification, and characterization of a haeminbinding lipoprotein from Haemophilus influenzae type b
    • Hanson, M.S., and Hansen, E.J. (1991) Molecular cloning, partial purification, and characterization of a haeminbinding lipoprotein from Haemophilus influenzae type b. Mol Microbiol 5: 267-278.
    • (1991) Mol Microbiol , vol.5 , pp. 267-278
    • Hanson, M.S.1    Hansen, E.J.2
  • 15
    • 0025105710 scopus 로고
    • Cloning and expression of a secondary Aspergillus niger pectin lyase gene (pelA): Indications of a pectin lyase gene family in A. niger
    • Harmsen, J.A.M., Kusters-van Someren, M.A., and Visser, J. (1990) Cloning and expression of a secondary Aspergillus niger pectin lyase gene (pelA): indications of a pectin lyase gene family in A. niger. Curr Genet 18: 161-166.
    • (1990) Curr Genet , vol.18 , pp. 161-166
    • Harmsen, J.A.M.1    Kusters-van Someren, M.A.2    Visser, J.3
  • 16
    • 0024853451 scopus 로고
    • Extracellular and periplasmic isoenzymes of pectate lyase from Erwinia carotovora subspecies carotovora belong to different gene family
    • Hinton, J.C.D., Sidebotham, J.M., Gill, D.R., and Salmond, G.P.C. (1989) Extracellular and periplasmic isoenzymes of pectate lyase from Erwinia carotovora subspecies carotovora belong to different gene family. Mol Microbiol 3: 1785-1795.
    • (1989) Mol Microbiol , vol.3 , pp. 1785-1795
    • Hinton, J.C.D.1    Sidebotham, J.M.2    Gill, D.R.3    Salmond, G.P.C.4
  • 17
    • 0024329301 scopus 로고
    • Isolation of Erwinia chrysanthemi mutants altered in pectinolytic enzyme production
    • Hugouvieux-Cotte-Pattat, N., and Robert-Baudouy, J. (1989) Isolation of Erwinia chrysanthemi mutants altered in pectinolytic enzyme production. Mol Microbiol 3: 1587-1597.
    • (1989) Mol Microbiol , vol.3 , pp. 1587-1597
    • Hugouvieux-Cotte-Pattat, N.1    Robert-Baudouy, J.2
  • 19
    • 0027258413 scopus 로고
    • Erwinia chrysanthemi EC16 produces a second set of plant-inducible pectate lyase isoenzymes
    • Kelemu, S., and Collmer, A. (1993) Erwinia chrysanthemi EC16 produces a second set of plant-inducible pectate lyase isoenzymes. Appl Env Microbiol 59: 1756-1761.
    • (1993) Appl Env Microbiol , vol.59 , pp. 1756-1761
    • Kelemu, S.1    Collmer, A.2
  • 20
    • 0026515440 scopus 로고
    • Listeria monocytogenes-induced actin assembly requires the actA gene product, a surface protein
    • Kocks, C., Gouin, E., Tabouret, M., Berche, P., Ohayon, H., and Cossart, P. (1992) Listeria monocytogenes-induced actin assembly requires the actA gene product, a surface protein. Cell 68: 521-531.
    • (1992) Cell , vol.68 , pp. 521-531
    • Kocks, C.1    Gouin, E.2    Tabouret, M.3    Berche, P.4    Ohayon, H.5    Cossart, P.6
  • 21
    • 0027172127 scopus 로고
    • Characterization and overexpression of the pem gene encoding pectin methylesterase of Erwinia chrysanthemi strain 3937
    • Laurent, F, Kotoujansky, A., Labesse, G., and Bertheau, Y. (1993) Characterization and overexpression of the pem gene encoding pectin methylesterase of Erwinia chrysanthemi strain 3937. Gene 131: 17-25.
    • (1993) Gene , vol.131 , pp. 17-25
    • Laurent, F.1    Kotoujansky, A.2    Labesse, G.3    Bertheau, Y.4
  • 23
    • 0025296153 scopus 로고
    • Molecular cloning and characterisation of an Erwinia carotovora subsp. carotovora pectin lyase gene that responds to DNA-damaging agents
    • McEvoy, J.L., Murata, H., and Chatterjee, A.K. (1990) Molecular cloning and characterisation of an Erwinia carotovora subsp. carotovora pectin lyase gene that responds to DNA-damaging agents. J Bacteriol 172: 3284-3289.
    • (1990) J Bacteriol , vol.172 , pp. 3284-3289
    • McEvoy, J.L.1    Murata, H.2    Chatterjee, A.K.3
  • 24
    • 0027336394 scopus 로고
    • A pH-independent assay for pectin methyl esterase for use in column chromatography
    • McMillan, G.P., Johnston, D.J., Morel, J-B., and Perombelon, M.C.M. (1993) A pH-independent assay for pectin methyl esterase for use in column chromatography. Anal Biochem 209: 377-379.
    • (1993) Anal Biochem , vol.209 , pp. 377-379
    • McMillan, G.P.1    Johnston, D.J.2    Morel, J.-B.3    Perombelon, M.C.M.4
  • 25
    • 0028106955 scopus 로고
    • An isoelectric focusing study of the effect of methyl-esterified pectic substances on the production of extracellular pectin isoenzymes by soft rot Erwinia spp
    • McMillan, M.P., Barrett, A.M., and Perombelon, M.C.M. (1994) An isoelectric focusing study of the effect of methyl-esterified pectic substances on the production of extracellular pectin isoenzymes by soft rot Erwinia spp. J Appl Bacteriol 77: 175-184.
    • (1994) J Appl Bacteriol , vol.77 , pp. 175-184
    • McMillan, M.P.1    Barrett, A.M.2    Perombelon, M.C.M.3
  • 26
    • 0023664635 scopus 로고
    • Sequence from picomole quantities of proteins electroblotted onto polyvinilidene difluoride membranes
    • Matsudaira, P. (1987) Sequence from picomole quantities of proteins electroblotted onto polyvinilidene difluoride membranes. J Biol Chem 262: 10035-10038.
    • (1987) J Biol Chem , vol.262 , pp. 10035-10038
    • Matsudaira, P.1
  • 27
    • 0003785155 scopus 로고
    • Cold Spring Harbour, New York: Cold Spring Harbour Laboratory Press
    • Miller, J. H. (1972) Experiments in Molecular Genetics. Cold Spring Harbour, New York: Cold Spring Harbour Laboratory Press.
    • (1972) Experiments in Molecular Genetics
    • Miller, J.H.1
  • 28
    • 0014252221 scopus 로고
    • Extracellular and intracellular polygalacturonic acid trans-eliminases of Erwinia carotovora
    • Moran, F., Nasuno, S., and Starr, M.P. (1968) Extracellular and intracellular polygalacturonic acid trans-eliminases of Erwinia carotovora. Arch Biochem Biophys 123: 293-306.
    • (1968) Arch Biochem Biophys , vol.123 , pp. 293-306
    • Moran, F.1    Nasuno, S.2    Starr, M.P.3
  • 29
    • 0028204482 scopus 로고
    • Specific interactions of the Erwinia chrysanthemi KdgR repressor with different operators of genes involved in pectinolysis
    • Nasser, W., Reverchon, S., Condemine, G., and Robert-Baudouy, J. (1994) Specific interactions of the Erwinia chrysanthemi KdgR repressor with different operators of genes involved in pectinolysis J Mol Biol 236: 427-440.
    • (1994) J Mol Biol , vol.236 , pp. 427-440
    • Nasser, W.1    Reverchon, S.2    Condemine, G.3    Robert-Baudouy, J.4
  • 30
    • 0028298380 scopus 로고
    • Isolation of outer membranes
    • Nikaido, H. (1994) Isolation of outer membranes. Meth Enzymol 235: 225-234.
    • (1994) Meth Enzymol , vol.235 , pp. 225-234
    • Nikaido, H.1
  • 31
    • 0016208980 scopus 로고
    • Inducibility of β-glucuronidase in wild-type and hexuronate-negative mutants of Escherichia coli K12
    • Novel, G., Didier-Fichet, M.-L., and Stoeber, F. (1974) Inducibility of β-glucuronidase in wild-type and hexuronate-negative mutants of Escherichia coli K12. J Bacteriol 102: 89-95.
    • (1974) J Bacteriol , vol.102 , pp. 89-95
    • Novel, G.1    Didier-Fichet, M.-L.2    Stoeber, F.3
  • 32
    • 0015327122 scopus 로고
    • Novel method for detection of β-lactamases by using a chromogenic cephalosporin substrate
    • O'Callaghan, C.H., Morris, A., Kirby, S.M., and Shingles, A.H. (1972) Novel method for detection of β-lactamases by using a chromogenic cephalosporin substrate. Antimicrob Ag Chemother 1: 283-288.
    • (1972) Antimicrob Ag Chemother , vol.1 , pp. 283-288
    • O'Callaghan, C.H.1    Morris, A.2    Kirby, S.M.3    Shingles, A.H.4
  • 33
    • 0015523228 scopus 로고
    • Mechanism of assembly of the outer membrane of Salmonella typhimurium
    • Osborn, M.J., Gander, J.E., Parisi, E., and Carson, J. (1972) Mechanism of assembly of the outer membrane of Salmonella typhimurium. J Biol Chem 247: 3962-3972.
    • (1972) J Biol Chem , vol.247 , pp. 3962-3972
    • Osborn, M.J.1    Gander, J.E.2    Parisi, E.3    Carson, J.4
  • 34
    • 0019435491 scopus 로고
    • Synthesis of the precursor to the B subunit of heat-labile enterotoxin in Escherichia coli
    • Palva, E.T., Hirst, T.R., Hardy, S.J.S., Holmgren, J., and Randall, L. (1981) Synthesis of the precursor to the B subunit of heat-labile enterotoxin in Escherichia coli. J Bacteriol 146: 325-330.
    • (1981) J Bacteriol , vol.146 , pp. 325-330
    • Palva, E.T.1    Hirst, T.R.2    Hardy, S.J.S.3    Holmgren, J.4    Randall, L.5
  • 35
    • 0023973502 scopus 로고
    • Molecular cloning and sequence of the pectin methyl esterase gene of Erwinia chrysanthemi B374
    • Plastow, G. S. (1988) Molecular cloning and sequence of the pectin methyl esterase gene of Erwinia chrysanthemi B374. Mol Microbiol 2: 247-254.
    • (1988) Mol Microbiol , vol.2 , pp. 247-254
    • Plastow, G.S.1
  • 36
    • 0028242862 scopus 로고
    • Molecular characterization of PulE, a protein required for pullulanase secretion
    • Possot, O., and Pugsley, A.P. (1994) Molecular characterization of PulE, a protein required for pullulanase secretion. Mol Microbiol 12: 287-299.
    • (1994) Mol Microbiol , vol.12 , pp. 287-299
    • Possot, O.1    Pugsley, A.P.2
  • 37
    • 0026599991 scopus 로고
    • Differential depolymerization mechanisms of pectate lyases secreted by Erwinia chrysanthemi EC16
    • Preston, J.F., Rice, J.D., Ingram, L.O., and Keen, N.T. (1992). Differential depolymerization mechanisms of pectate lyases secreted by Erwinia chrysanthemi EC16. J Bacteriol 174: 2039-2042.
    • (1992) J Bacteriol , vol.174 , pp. 2039-2042
    • Preston, J.F.1    Rice, J.D.2    Ingram, L.O.3    Keen, N.T.4
  • 38
    • 0027450561 scopus 로고
    • The complete general secretory pathway in Gram-negative bacteria
    • Pugsley, A.P. (1993) The complete general secretory pathway in Gram-negative bacteria. Microbiol Rev 57: 50-108.
    • (1993) Microbiol Rev , vol.57 , pp. 50-108
    • Pugsley, A.P.1
  • 39
    • 0022633064 scopus 로고
    • Extracellular pullulanase of Klebsiella pneumoniae is a lipoprotein
    • Pugsley, A.P., Chapon, C., and Schwartz, M. (1986) Extracellular pullulanase of Klebsiella pneumoniae is a lipoprotein. J Bacteriol 166: 1083-1088.
    • (1986) J Bacteriol , vol.166 , pp. 1083-1088
    • Pugsley, A.P.1    Chapon, C.2    Schwartz, M.3
  • 40
    • 0021179654 scopus 로고
    • Phi-EC2, a new generalized transducing phage of Erwinia chrysanthemi
    • Résibois, A., Colet, M., Faelen, M., Schoonejans, E., and Toussaint, A. (1984) Phi-EC2, a new generalized transducing phage of Erwinia chrysanthemi. Virology 137: 102-112.
    • (1984) Virology , vol.137 , pp. 102-112
    • Résibois, A.1    Colet, M.2    Faelen, M.3    Schoonejans, E.4    Toussaint, A.5
  • 41
    • 0025761081 scopus 로고
    • Characterization of kdgR, a gene of Erwinia chrysanthemi that regulates pectin degradation
    • Reverchon, S., Nasser., W., and Robert-Baudouy, J. (1991) Characterization of kdgR, a gene of Erwinia chrysanthemi that regulates pectin degradation. Mol Microbiol 5: 2203-2216.
    • (1991) Mol Microbiol , vol.5 , pp. 2203-2216
    • Reverchon, S.1    Nasser, W.2    Robert-Baudouy, J.3
  • 42
    • 0028334118 scopus 로고
    • pecS: A locus controlling pectinase, cellulase and blue pigment production in Erwinia chrysanthemi
    • Reverchon, S., Nasser, W., and Robert-Baudouy, J. (1994) pecS: a locus controlling pectinase, cellulase and blue pigment production in Erwinia chrysanthemi. Mol Microbiol 11: 1127-1139.
    • (1994) Mol Microbiol , vol.11 , pp. 1127-1139
    • Reverchon, S.1    Nasser, W.2    Robert-Baudouy, J.3
  • 43
    • 0022344451 scopus 로고
    • Marker exchange mutagenesis of a pectate lyase isoenzyme gene in Erwinia chrysanthemi
    • Roeder, D.L., and Collmer, A. (1985). Marker exchange mutagenesis of a pectate lyase isoenzyme gene in Erwinia chrysanthemi. J Bacteriol 164: 51-56.
    • (1985) J Bacteriol , vol.164 , pp. 51-56
    • Roeder, D.L.1    Collmer, A.2
  • 46
    • 0023049475 scopus 로고
    • Purification and properties of membrane-bound hydrogenase isoenzyme 1 from anaerobically grown Escherichia coli K12
    • Sawers, R.G., and Boxer, D.H. (1986) Purification and properties of membrane-bound hydrogenase isoenzyme 1 from anaerobically grown Escherichia coli K12. Eur J Biochem 156: 265-275.
    • (1986) Eur J Biochem , vol.156 , pp. 265-275
    • Sawers, R.G.1    Boxer, D.H.2
  • 47
    • 0015132002 scopus 로고
    • Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100
    • Schnaitman, C.A. (1971) Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. J Bacteriol 108: 545-552.
    • (1971) J Bacteriol , vol.108 , pp. 545-552
    • Schnaitman, C.A.1
  • 48
    • 0023761294 scopus 로고
    • Isoenzymes of extracellular pectate lyases of bacteria of the genus Erwinia
    • Russ
    • Shevchik, V.E., Evtushenkov, A.N., and Fomichev, Y.K. (1988) Isoenzymes of extracellular pectate lyases of bacteria of the genus Erwinia. Biokhimiya (Russ) 53: 1628-1638.
    • (1988) Biokhimiya , vol.53 , pp. 1628-1638
    • Shevchik, V.E.1    Evtushenkov, A.N.2    Fomichev, Y.K.3
  • 49
    • 0026087795 scopus 로고
    • Molecular cloning and sequencing of a pectinesterase gene from Pseudomonas solanacearum
    • Spök, A., Schoengendorfer, K., and Schwab, H. (1991) Molecular cloning and sequencing of a pectinesterase gene from Pseudomonas solanacearum. J Gen Microbiol 137: 131-140.
    • (1991) J Gen Microbiol , vol.137 , pp. 131-140
    • Spök, A.1    Schoengendorfer, K.2    Schwab, H.3
  • 50
    • 0021919826 scopus 로고
    • A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes
    • Tabor, S., and Richardson, C. (1985) A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci USA 82: 1074-1078.
    • (1985) Proc Natl Acad Sci USA , vol.82 , pp. 1074-1078
    • Tabor, S.1    Richardson, C.2
  • 51
    • 0014690791 scopus 로고
    • The reliability of molecular weight determination by dodecyl sulphate-polyacrylamide gel electrophoresis
    • Weber, K., and Osborn, M. (1969) The reliability of molecular weight determination by dodecyl sulphate-polyacrylamide gel electrophoresis. J Biol Chem 244: 4406-4412.
    • (1969) J Biol Chem , vol.244 , pp. 4406-4412
    • Weber, K.1    Osborn, M.2
  • 52
    • 0024279918 scopus 로고
    • A single amino acid determinant of the membrane localization of lipoproteins in E. coli
    • Yamaguchi, K., Wu, F., and Inouye, M. (1988) A single amino acid determinant of the membrane localization of lipoproteins in E. coli. Cell 53: 423-432.
    • (1988) Cell , vol.53 , pp. 423-432
    • Yamaguchi, K.1    Wu, F.2    Inouye, M.3


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.