Taoka Azuma, Asada Ryuji, Sasaki Hideaki, Anzawa Kazushi, Wu Long-Fei, Fukumori Yoshihiro
Department of Life Science, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
J Bacteriol. 2006 Jun;188(11):3805-12. doi: 10.1128/JB.00020-06.
Magnetospirillum magnetotacticum possesses intracellular magnetite particles with a chain-like structure, termed magnetosomes. The bacterium expresses 22-kDa and 12-kDa magnetosome-associated proteins, termed Mam22 (MamA) and Mam12 (MamC), respectively. In this study, we investigated the structure of the purified magnetosomes with transmission electron microscopic techniques and found that the magnetosomes consisted of four compartments, i.e., magnetite crystal, magnetosomal membrane, interparticle connection, and magnetosomal matrix. Furthermore, we determined the precise localizations of Mam22 and Mam12 using immunogold staining of the purified magnetosomes and ultrathin sections of the bacterial cells. Interestingly, most Mam22 existed in the magnetosomal matrix, whereas Mam12 was strictly localized in the magnetosomal membrane. Moreover, the recombinant Mam22 was attached to the magnetosomal matrix of the Mam22-deficient magnetosomes prepared by alkaline treatment, such as 0.1 M Caps-NaOH buffer (pH 11.0). The spatial localization of the magnetosome-associated proteins in the magnetosomal chain provides useful information to elucidate the functional roles of these proteins.
趋磁螺菌拥有细胞内具有链状结构的磁铁矿颗粒,称为磁小体。该细菌表达分别称为Mam22(MamA)和Mam12(MamC)的22 kDa和12 kDa磁小体相关蛋白。在本研究中,我们用透射电子显微镜技术研究了纯化磁小体的结构,发现磁小体由四个部分组成,即磁铁矿晶体、磁小体膜、颗粒间连接和磁小体基质。此外,我们通过对纯化磁小体和细菌细胞超薄切片进行免疫金染色,确定了Mam22和Mam12的精确定位。有趣的是,大多数Mam22存在于磁小体基质中,而Mam12严格定位于磁小体膜中。此外,重组Mam22附着于通过碱性处理(如0.1 M Caps-NaOH缓冲液(pH 11.0))制备的Mam22缺陷型磁小体的磁小体基质上。磁小体相关蛋白在磁小体链中的空间定位为阐明这些蛋白的功能作用提供了有用信息。