Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented "EF-Hand-Like" Calcium-Binding Phage Lysin

CORC > 青岛生物能源与过程研究所 > 中国科学院青岛生物能源与过程研究所 > 代谢物组学团队

	Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented "EF-Hand-Like" Calcium-Binding Phage Lysin
	Gu, Jingmin 1; Feng, Yingang 2; Feng, Xin 1; Sun, Changjiang 1; Lei, Liancheng 1; Ding, Wei 3; Niu, Fengfeng 4; Jiao, Lianying 4; Yang, Mei 1; Li, Yue 1
刊名	PLOS PATHOGENS
	2014-05-01
卷号	10 期号:5
英文摘要	The lysin LysGH15, which is derived from the staphylococcal phage GH15, demonstrates a wide lytic spectrum and strong lytic activity against methicillin-resistant Staphylococcus aureus (MRSA). Here, we find that the lytic activity of the full-length LysGH15 and its CHAP domain is dependent on calcium ions. To elucidate the molecular mechanism, the structures of three individual domains of LysGH15 were determined. Unexpectedly, the crystal structure of the LysGH15 CHAP domain reveals an "EF-hand-like" calcium-binding site near the Cys-His-Glu-Asn quartet active site groove. To date, the calcium-binding site in the LysGH15 CHAP domain is unique among homologous proteins, and it represents the first reported calcium-binding site in the CHAP family. More importantly, the calcium ion plays an important role as a switch that modulates the CHAP domain between the active and inactive states. Structure-guided mutagenesis of the amidase-2 domain reveals that both the zinc ion and E282 are required in catalysis and enable us to propose a catalytic mechanism. Nuclear magnetic resonance (NMR) spectroscopy and titration-guided mutagenesis identify residues (e. g., N404, Y406, G407, and T408) in the SH3b domain that are involved in the interactions with the substrate. To the best of our knowledge, our results constitute the first structural information on the biochemical features of a staphylococcal phage lysin and represent a pivotal step forward in understanding this type of lysin.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Microbiology ; Parasitology ; Virology
研究领域[WOS]	Microbiology ; Parasitology ; Virology
关键词[WOS]	RESISTANT STAPHYLOCOCCUS-AUREUS ; MACROMOLECULAR STRUCTURES ; BACTERIOPHAGE LYSIN ; CRYSTAL-STRUCTURE ; LYTIC ACTIVITY ; STREPTOCOCCUS-AGALACTIAE ; PROPHAGE-ENDOLYSIN ; PROTEIN STRUCTURES ; TARGETING DOMAIN ; NOE ASSIGNMENT
收录类别	SCI
语种	英语
WOS记录号	WOS:000337732300026
公开日期	2015-12-24
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn/handle/337004/6255]
专题	青岛生物能源与过程研究所_代谢物组学团队
作者单位	1.Jilin Univ, Coll Vet Med, Minist Educ, Key Lab Zoonosis, Changchun 130023, Peoples R China 2.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Energy Genet, Qingdao, Peoples R China 3.Chinese Acad Sci, Inst Biophys, Ctr Biol Imaging, Beijing 100080, Peoples R China 4.Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100080, Peoples R China
推荐引用方式 GB/T 7714	Gu, Jingmin,Feng, Yingang,Feng, Xin,et al. Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented "EF-Hand-Like" Calcium-Binding Phage Lysin[J]. PLOS PATHOGENS,2014,10(5).
APA	Gu, Jingmin.,Feng, Yingang.,Feng, Xin.,Sun, Changjiang.,Lei, Liancheng.,...&Han, Wenyu.(2014).Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented "EF-Hand-Like" Calcium-Binding Phage Lysin.PLOS PATHOGENS,10(5).
MLA	Gu, Jingmin,et al."Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented "EF-Hand-Like" Calcium-Binding Phage Lysin".PLOS PATHOGENS 10.5(2014).