Trost J T, Brune D C, Blankenship R E
Department of Chemistry and Biochemistry, Arizona State University, Tempe 85287-1604, USA.
Photosynth Res. 1992;32:11-22.
Photosynthetic reaction centers isolated from Heliobacillus mobilis exhibit a single major protein on SDS-PAGE of 47 000 Mr. Attempts to sequence the reaction center polypeptide indicated that the N-terminus is blocked. After enzymatic and chemical cleavage, four peptide fragments were sequenced from the Heliobacillus mobilis apoprotein. Only one of these sequences showed significant specific similarity to any of the protein and deduced protein sequences in the GenBank data base. This fragment is identical with 56% of the residues, including both cysteines, found in highly conserved region that is proposed to bind iron-sulfur center Fx in the Photosystem I reaction center peptide that is the psaB gene product. The similarity to the psaA gene product in this region is 48%. Redox titrations of laser-flash-induced photobleaching with millisecond decay kinetics on isolated reaction centers from Heliobacterium gestii indicate a midpoint potential of -414 mV with n = 2 titration behavior. In membranes, the behavior is intermediate between n = 1 and n = 2, and the apparent midpoint potential is -444 mV. This is compared to the behavior in Photosystem I, where the intermediate electron acceptor A1, thought to be a phylloquinone molecule, has been proposed to undergo a double reduction at low redox potentials in the presence of viologen redox mediators. These results strongly suggest that the acceptor side electron transfer system in reaction centers from heliobacteria is indeed analogous to that found in Photosystem I. The sequence similarities indicate that the divergence of the heliobacteria from the Photosystem I line occurred before the gene duplication and subsequent divergence that lead to the heterodimeric protein core of the Photosystem I reaction center.
从运动太阳杆菌中分离出的光合反应中心在SDS-PAGE上显示出一条分子量为47000的主要蛋白质条带。对反应中心多肽进行测序的尝试表明其N端被封闭。经过酶切和化学裂解后,从运动太阳杆菌脱辅基蛋白中获得了四个肽段并进行了测序。这些序列中只有一个与GenBank数据库中的任何蛋白质及推导的蛋白质序列显示出显著的特异性相似性。该片段与光系统I反应中心肽(即psaB基因产物)中拟结合铁硫中心Fx的高度保守区域中的56%的残基相同,包括两个半胱氨酸。该区域与psaA基因产物的相似性为48%。对来自Gestii太阳杆菌的分离反应中心进行的具有毫秒级衰减动力学的激光闪光诱导光漂白的氧化还原滴定表明,中点电位为-414 mV,滴定行为为n = 2。在膜中,行为介于n = 1和n = 2之间,表观中点电位为-444 mV。相比之下,在光系统I中,中间电子受体A1被认为是一个叶醌分子,有人提出在存在紫精氧化还原介质的情况下,它在低氧化还原电位下会发生双还原。这些结果强烈表明,太阳杆菌反应中心的受体侧电子传递系统确实与光系统I中的类似。序列相似性表明,太阳杆菌与光系统I谱系的分化发生在导致光系统I反应中心异二聚体蛋白质核心的基因复制及随后的分化之前。