Open AccessCrystallization and preliminary X‐ray diffraction analysis of l , l ‐diaminopimelate aminotransferase (DapL) from Chlamydomonas reinhardtii
Author(s) -
Hudson André O.,
Girón Irma,
Dobson Renwick C. J.
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430911004844x
Subject(s) - chlamydomonas reinhardtii , crystallization , chemistry , resolution (logic) , crystallography , recombinant dna , bacteria , lysine , biochemistry , stereochemistry , amino acid , biology , gene , genetics , organic chemistry , artificial intelligence , computer science , mutant
In the anabolic synthesis of diaminopimelate and lysine in plants and in some bacteria, the enzyme l , l ‐diaminopimelate aminotransferase (DapL; EC 2.6.1.83) catalyzes the conversion of tetrahydrodipicolinic acid (THDPA) to l , l ‐diaminopimelate, bypassing the DapD, DapC and DapE enzymatic steps in the bacterial acyl pathways. Here, the cloning, expression, purification, crystallization and preliminary X‐ray diffraction analysis of DapL from the alga Chlamydomonas reinhardtii are presented. Protein crystals were grown in conditions containing 25%( w / v ) PEG 3350 and 200 m M lithium sulfate and initially diffracted to ∼1.35 Å resolution. They belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 58.9, b = 91.8, c = 162.9 Å. The data were processed to 1.55 Å resolution with an R merge of 0.081, an R p.i.m. of 0.044, an R r.i.m of 0.093 and a V M of 2.28 Å 3 Da −1 .