Cloning, expression, crystallization and preliminary crystallographic analysis of a pentapeptide-repeat protein (Rfr23) from the bacterium Cyanothece 51142. Export

@article{citeulike:971178, abstract = {A unique feature of cyanobacteria genomes is the abundance of genes that code for hypothetical proteins containing tandem pentapeptide repeats approximately described by the consensus motif A(N/D)LXX. To date, the structures of two pentapeptide-repeat proteins (PRPs) have been determined, with the tandem pentapeptide-repeat sequences observed to adopt a novel type of right-handed quadrilateral beta-helix, or Rfr-fold, in both structures. One structure, Mycobacterium tuberculosis MfpA, is a 183-residue protein that contains 30 consecutive pentapeptide repeats and appears to offer antibiotic resistance by acting as a DNA mimic. The other structure, Cyanothece 51142 Rfr32, is a 167-residue protein that contains 21 consecutive pentapeptide repeats. The function of Rfr32, like the other 35 hypothetical PRPs identified in the genome of Cyanothece, is unknown. In an effort to understand the role of PRPs in cyanobacteria and to better characterize the structural properties of Rfr-folds with different amino-acid sequences, a second PRP from Cyanothece 51142, Rfr23, has been cloned, expressed and purified. Selenomethione-substituted protein was crystallized by vapor diffusion in hanging drops. Nearly complete SAD and native diffraction data sets were collected from these crystals to 2.5 and 2.1 A resolution, respectively, using synchrotron radiation. The crystals belonged to space group I4(1), with unit-cell parameters a = b = 106.61, c = 53.37 A, and one molecule per asymmetric unit. Preliminary analysis of the electron-density map from the SAD data shows that Rfr23 contains an Rfr-fold.}, author = {Buchko, Garry W. and Robinson, Howard and Ni, Shuisong and Pakrasi, Himadri B. and Kennedy, Michael A.}, citeulike-article-id = {971178}, citeulike-linkout-0 = {http://dx.doi.org/10.1107/S174430910604663X}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/17142909}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=17142909}, day = {1}, doi = {10.1107/S174430910604663X}, issn = {1744-3091}, journal = {Acta crystallographica. Section F, Structural biology and crystallization communications}, keywords = {beta\_helix, beta\_solenoid}, month = {December}, number = {Pt 12}, pages = {1251--1254}, posted-at = {2009-09-30 12:19:52}, priority = {2}, publisher = {International Union of Crystallography}, title = {Cloning, expression, crystallization and preliminary crystallographic analysis of a pentapeptide-repeat protein (Rfr23) from the bacterium Cyanothece 51142.}, url = {http://dx.doi.org/10.1107/S174430910604663X}, volume = {62}, year = {2006} }

TY - JOUR ID - citeulike:971178 L3 - citeulike-article-id:971178 N2 - A unique feature of cyanobacteria genomes is the abundance of genes that code for hypothetical proteins containing tandem pentapeptide repeats approximately described by the consensus motif A(N/D)LXX. To date, the structures of two pentapeptide-repeat proteins (PRPs) have been determined, with the tandem pentapeptide-repeat sequences observed to adopt a novel type of right-handed quadrilateral beta-helix, or Rfr-fold, in both structures. One structure, Mycobacterium tuberculosis MfpA, is a 183-residue protein that contains 30 consecutive pentapeptide repeats and appears to offer antibiotic resistance by acting as a DNA mimic. The other structure, Cyanothece 51142 Rfr32, is a 167-residue protein that contains 21 consecutive pentapeptide repeats. The function of Rfr32, like the other 35 hypothetical PRPs identified in the genome of Cyanothece, is unknown. In an effort to understand the role of PRPs in cyanobacteria and to better characterize the structural properties of Rfr-folds with different amino-acid sequences, a second PRP from Cyanothece 51142, Rfr23, has been cloned, expressed and purified. Selenomethione-substituted protein was crystallized by vapor diffusion in hanging drops. Nearly complete SAD and native diffraction data sets were collected from these crystals to 2.5 and 2.1 A resolution, respectively, using synchrotron radiation. The crystals belonged to space group I4(1), with unit-cell parameters a = b = 106.61, c = 53.37 A, and one molecule per asymmetric unit. Preliminary analysis of the electron-density map from the SAD data shows that Rfr23 contains an Rfr-fold. IS - Pt 12 JF - Acta crystallographica. Section F, Structural biology and crystallization communications SN - 1744-3091 EP - 1254 TI - Cloning, expression, crystallization and preliminary crystallographic analysis of a pentapeptide-repeat protein (Rfr23) from the bacterium Cyanothece 51142. PB - International Union of Crystallography VL - 62 SP - 1251 KW - beta_helix KW - beta_solenoid AU - Buchko, Garry AU - Robinson, Howard AU - Ni, Shuisong AU - Pakrasi, Himadri AU - Kennedy, Michael PY - 2006/12/01/ UR - http://dx.doi.org/10.1107/S174430910604663X ER -