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

doi:10.1371/journal.ppat.1004109

QIBEBT-IR > 代谢物组学研究组

	Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented "EF-Hand-Like" Calcium-Binding Phage Lysin
	Gu, Jingmin 1; Feng, Yingang2 ; Feng, Xin 1; Sun, Changjiang 1; Lei, Liancheng 1; Ding, Wei 3; Niu, Fengfeng 4; Jiao, Lianying 4; Yang, Mei 1; Li, Yue 1; Liu, Xiaohe 1; Song, Jun 1; Cui, Ziyin 1; Han, Dong 1; Du, Chongtao 1; Yang, Yongjun 1; Ouyang, Songying 4; Liu, Zhi-Jie 4; Han, Wenyu1
	2014-05-01
发表期刊	PLOS PATHOGENS
卷号	10 期号:5
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.
文章类型	Article
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
DOI	10.1371/journal.ppat.1004109
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研究方向	Microbiology ; Parasitology ; Virology
WOS类目	Microbiology ; Parasitology ; Virology
WOS记录号	WOS:000337732300026
引用统计
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/6255
专题	代谢物组学研究组
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).