Open AccessCrystallization and X‐ray diffraction studies of crustacean proliferating cell nuclear antigen
Author(s) -
CarrascoMiranda Jesus S.,
CardonaFelix Cesar S.,
LopezZavala Alonso A.,
delaReVega Enrique,
De la Mora Eugenio,
RudiñoPiñera Enrique,
SoteloMundo Rogerio R.,
Brieba Luis G.
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112040444
Subject(s) - proliferating cell nuclear antigen , biology , antigen , dna replication , crystallization , dna , recombinant dna , nuclear protein , microbiology and biotechnology , crystallography , biophysics , biochemistry , chemistry , gene , genetics , transcription factor , organic chemistry
Proliferating cell nuclear antigen (PCNA), a member of the sliding clamp family of proteins, interacts specifically with DNA replication and repair proteins through a small peptide motif called the PCNA‐interacting protein or PIP box. PCNA is recognized as one of the key proteins involved in DNA metabolism. In the present study, the recombinant PCNA from Litopenaeus vannamei ( Lv PCNA) was heterologously overexpressed and purified using metal ion‐affinity chromatography. Crystals suitable for diffraction grew overnight using the hanging‐drop vapour‐diffusion method. Lv PCNA crystals belong to space group C 2 with unit‐cell parameters a = 144.6, b = 83.4, c = 74.3 Å, β = 117.6°. One data set was processed to 3 Å resolution, with an overall R meas of 0.09 and a completeness of 93.3%. Initial phases were obtained by molecular replacement using a homology model of Lv PCNA as the search model. Refinement and structural analysis are underway. This report is the first successful crystallographic analysis of a marine crustacean decapod shrimp ( L. vannamei ) proliferating cell nuclear antigen.