Gardberg Anna, Abendroth Jan, Bhandari Janhavi, Sankaran Banumathi, Staker Bart
Emerald BioStructures, 7869 NE Day Road West, Bainbridge Island, WA 98110, USA.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Sep 1;67(Pt 9):1051-4. doi: 10.1107/S174430911101894X. Epub 2011 Aug 13.
Fructose bisphosphate aldolase (FBPA) enzymes have been found in a broad range of eukaryotic and prokaryotic organisms. FBPA catalyses the cleavage of fructose 1,6-bisphosphate into glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. The SSGCID has reported several FBPA structures from pathogenic sources, including the bacterium Brucella melitensis and the protozoan Babesia bovis. Bioinformatic analysis of the Bartonella henselae genome revealed an FBPA homolog. The B. henselae FBPA enzyme was recombinantly expressed and purified for X-ray crystallographic studies. The purified enzyme crystallized in the apo form but failed to diffract; however, well diffracting crystals could be obtained by cocrystallization in the presence of the native substrate fructose 1,6-bisphosphate. A data set to 2.35 Å resolution was collected from a single crystal at 100 K. The crystal belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a=72.39, b=127.71, c=157.63 Å. The structure was refined to a final free R factor of 22.2%. The structure shares the typical barrel tertiary structure and tetrameric quaternary structure reported for previous FBPA structures and exhibits the same Schiff base in the active site.
在广泛的真核生物和原核生物中都发现了果糖二磷酸醛缩酶（FBPA）。FBPA催化1,6-二磷酸果糖裂解为3-磷酸甘油醛和磷酸二羟丙酮。SSGCID报道了几种来自致病源的FBPA结构，包括细菌布鲁氏菌和原生动物牛巴贝斯虫。对汉赛巴尔通体基因组的生物信息学分析揭示了一种FBPA同源物。重组表达并纯化了汉赛巴尔通体FBPA酶，用于X射线晶体学研究。纯化后的酶以脱辅基形式结晶，但无法衍射；然而，在天然底物1,6-二磷酸果糖存在下共结晶可以获得良好衍射的晶体。在100 K下从单晶收集了分辨率为2.35 Å的数据集。该晶体属于正交空间群P2(1)2(1)2(1)，晶胞参数a = 72.39，b = 127.71，c = 157.63 Å。结构精修后的最终自由R因子为22.2%。该结构具有先前报道的FBPA结构所具有的典型桶状三级结构和四聚体四级结构，并且在活性位点呈现相同的席夫碱。
