Buchko Garry W, Abendroth Jan, Clifton Matthew C, Robinson Howard, Zhang Yanfeng, Hewitt Stephen N, Staker Bart L, Edwards Thomas E, Van Voorhis Wesley C, Myler Peter J
Seattle Structural Genomics Center for Infectious Disease, USA.
Biology Department, Brookhaven National Laboratory, Upton, New York, USA.
Acta Crystallogr F Struct Biol Commun. 2015 May;71(Pt 5):522-30. doi: 10.1107/S2053230X14028210. Epub 2015 Apr 18.
Cryptosporidiosis is an infectious disease caused by protozoan parasites of the Cryptosporidium genus. Infection is associated with mild to severe diarrhea that usually resolves spontaneously in healthy human adults, but may lead to severe complications in young children and in immunocompromised patients. The genome of C. parvum contains a gene, CUTA_CRYPI, that may play a role in regulating the intracellular concentration of copper, which is a toxic element in excess. Here, the crystal structure of this CutA1 protein, Cp-CutA1, is reported at 2.0 Å resolution. As observed for other CutA1 structures, the 117-residue protein is a trimer with a core ferrodoxin-like fold. Circular dichroism spectroscopy shows little, in any, unfolding of Cp-CutA1 up to 353 K. This robustness is corroborated by (1)H-(15)N HSQC spectra at 333 K, which are characteristic of a folded protein, suggesting that NMR spectroscopy may be a useful tool to further probe the function of the CutA1 proteins. While robust, Cp-CutA1 is not as stable as the homologous protein from a hyperthermophile, perhaps owing to a wide β-bulge in β2 that protrudes Pro48 and Ser49 outside the β-sheet.
隐孢子虫病是一种由隐孢子虫属原生动物寄生虫引起的传染病。感染与轻度至重度腹泻有关,在健康成年人中通常会自发缓解,但在幼儿和免疫功能低下的患者中可能会导致严重并发症。微小隐孢子虫的基因组包含一个基因CUTA_CRYPI,该基因可能在调节铜的细胞内浓度方面发挥作用,过量的铜是一种有毒元素。在此,报道了这种CutA1蛋白Cp-CutA1在2.0 Å分辨率下的晶体结构。正如在其他CutA1结构中观察到的那样,这种由117个残基组成的蛋白质是一个三聚体,具有类似铁氧化还原蛋白的核心折叠结构。圆二色光谱显示,直到353 K,Cp-CutA1几乎没有展开。333 K下的(1)H-(15)N HSQC光谱证实了这种稳定性,该光谱是折叠蛋白的特征,表明核磁共振光谱可能是进一步探究CutA1蛋白功能的有用工具。虽然Cp-CutA1很稳定,但它不如来自嗜热菌的同源蛋白稳定,这可能是由于β2中的一个宽β凸起将Pro48和Ser49突出到β折叠之外。