title: Binding of cyclic diguanylate in the non-catalytic EAL domain of FimX induces a long-range conformational change.
abstract: FimX is a multidomain signaling protein required for type IV pilus biogenesis and twitching motility in the opportunistic pathogen Pseudomonas aeruginosa. FimX is localized to the single pole of the bacterial cell, and the unipolar localization is crucial for the correct assembly of type IV pili. FimX contains a non-catalytic EAL domain that lacks cyclic diguanylate (c-di-GMP) phosphodiesterase activity. It was shown that deletion of the EAL domain or mutation of the signature EVL motif affects the unipolar localization of FimX. However, it was not understood how the C-terminal EAL domain could influence protein localization considering that the localization sequence resides in the remote N-terminal region of the protein. Using hydrogen/deuterium exchange-coupled mass spectrometry, we found that the binding of c-di-GMP to the EAL domain triggers a long-range (∼ca. 70 Å) conformational change in the N-terminal REC domain and the adjacent linker. In conjunction with the observation that mutation of the EVL motif of the EAL domain abolishes the binding of c-di-GMP, the hydrogen/deuterium exchange results provide a molecular explanation for the mediation of protein localization and type IV pilus biogenesis by c-di-GMP through a remarkable allosteric regulation mechanism.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore 637551.
authors.forename: Yaning
authors.initials: Y
authors.lastname: Qi
authors.forename: Mary Lay Cheng
authors.initials: ML
authors.lastname: Chuah
authors.forename: Xueming
authors.initials: X
authors.lastname: Dong
authors.forename: Kailing
authors.initials: K
authors.lastname: Xie
authors.forename: Zhen
authors.initials: Z
authors.lastname: Luo
authors.forename: Kai
authors.initials: K
authors.lastname: Tang
authors.forename: Zhao-Xun
authors.initials: ZX
authors.lastname: Liang
mesh_terms: D001057:Apolipoproteins E / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D016174:Hepacivirus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D013058:Mass Spectrometry; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D001057:Apolipoproteins E; D008563:Membrane Lipids; D014759:Viral Envelope Proteins