Zubieta Chloe, Krishna S Sri, Kapoor Mili, Kozbial Piotr, McMullan Daniel, Axelrod Herbert L, Miller Mitchell D, Abdubek Polat, Ambing Eileen, Astakhova Tamara, Carlton Dennis, Chiu Hsiu-Ju, Clayton Thomas, Deller Marc C, Duan Lian, Elsliger Marc-André, Feuerhelm Julie, Grzechnik Slawomir K, Hale Joanna, Hampton Eric, Han Gye Won, Jaroszewski Lukasz, Jin Kevin K, Klock Heath E, Knuth Mark W, Kumar Abhinav, Marciano David, Morse Andrew T, Nigoghossian Edward, Okach Linda, Oommachen Silvya, Reyes Ron, Rife Christopher L, Schimmel Paul, van den Bedem Henry, Weekes Dana, White Aprilfawn, Xu Qingping, Hodgson Keith O, Wooley John, Deacon Ashley M, Godzik Adam, Lesley Scott A, Wilson Ian A
Stanford Synchrotron Radiation Laboratory, Stanford University, Menlo Park, California, USA.
Proteins. 2007 Nov 1;69(2):223-33. doi: 10.1002/prot.21550.
BtDyP from Bacteroides thetaiotaomicron (strain VPI-5482) and TyrA from Shewanella oneidensis are dye-decolorizing peroxidases (DyPs), members of a new family of heme-dependent peroxidases recently identified in fungi and bacteria. Here, we report the crystal structures of BtDyP and TyrA at 1.6 and 2.7 A, respectively. BtDyP assembles into a hexamer, while TyrA assembles into a dimer; the dimerization interface is conserved between the two proteins. Each monomer exhibits a two-domain, alpha+beta ferredoxin-like fold. A site for heme binding was identified computationally, and modeling of a heme into the proposed active site allowed for identification of residues likely to be functionally important. Structural and sequence comparisons with other DyPs demonstrate a conservation of putative heme-binding residues, including an absolutely conserved histidine. Isothermal titration calorimetry experiments confirm heme binding, but with a stoichiometry of 0.3:1 (heme:protein).
来自嗜热栖热放线菌（菌株VPI-5482）的BtDyP和来自腐败希瓦氏菌的TyrA是染料脱色过氧化物酶（DyPs），它们是最近在真菌和细菌中发现的一个新的血红素依赖性过氧化物酶家族的成员。在此，我们分别报道了BtDyP和TyrA在1.6 Å和2.7 Å分辨率下的晶体结构。BtDyP组装成六聚体，而TyrA组装成二聚体；两种蛋白质之间的二聚化界面是保守的。每个单体呈现出一种双结构域的α+β铁氧化还原蛋白样折叠。通过计算确定了血红素结合位点，并将血红素建模到提议的活性位点中，从而能够鉴定可能具有功能重要性的残基。与其他DyPs的结构和序列比较表明，假定的血红素结合残基具有保守性，包括一个绝对保守的组氨酸。等温滴定量热实验证实了血红素结合，但化学计量比为0.3:1（血红素:蛋白质）。
