5EOU

Pseudomonas aeruginosa PilM:PilN1-12 bound to ATP

Summary for 5EOU

• Entry DOI: 10.2210/pdb5eou/pdb
• Related: 5EOX 5EOY
• Descriptor: Type 4 fimbrial biogenesis protein PilM,Type 4 fimbrial biogenesis protein PilN, SULFATE ION, CHLORIDE ION, ... (7 entities in total)
• Functional Keywords: pilm, piln, type iv pilus, t4p, peptide binding protein
• Biological source: Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228); More
• Total number of polymer chains: 2
• Total formula weight: 81601.24

Authors

McCallum, M.,Tammam, S.,Robinson, H.,Shah, M.,Calmettes, C.,Moraes, T.,Burrows, L.,Howell, L.P. (deposition date: 2015-11-10, release date: 2016-04-27, Last modification date: 2023-09-27)

Primary citation

McCallum, M.,Tammam, S.,Little, D.J.,Robinson, H.,Koo, J.,Shah, M.,Calmettes, C.,Moraes, T.F.,Burrows, L.L.,Howell, P.L.
PilN Binding Modulates the Structure and Binding Partners of the Pseudomonas aeruginosa Type IVa Pilus Protein PilM.
J.Biol.Chem., 291:11003-11015, 2016

PubMed Abstract: Pseudomonas aeruginosa is an opportunistic bacterial pathogen that expresses type IVa pili. The pilus assembly system, which promotes surface-associated twitching motility and virulence, is composed of inner and outer membrane subcomplexes, connected by an alignment subcomplex composed of PilMNOP. PilM binds to the N terminus of PilN, and we hypothesize that this interaction causes functionally significant structural changes in PilM. To characterize this interaction, we determined the crystal structures of PilM and a PilM chimera where PilM was fused to the first 12 residues of PilN (PilM·PilN(1-12)). Structural analysis, multiangle light scattering coupled with size exclusion chromatography, and bacterial two-hybrid data revealed that PilM forms dimers mediated by the binding of a novel conserved motif in the N terminus of PilM, and binding PilN abrogates this binding interface, resulting in PilM monomerization. Structural comparison of PilM with PilM·PilN(1-12) revealed that upon PilN binding, there is a large domain closure in PilM that alters its ATP binding site. Using biolayer interferometry, we found that the association rate of PilN with PilM is higher in the presence of ATP compared with ADP. Bacterial two-hybrid data suggested the connectivity of the cytoplasmic and inner membrane components of the type IVa pilus machinery in P. aeruginosa, with PilM binding to PilB, PilT, and PilC in addition to PilN. Pull-down experiments demonstrated direct interactions of PilM with PilB and PilT. We propose a working model in which dynamic binding of PilN facilitates functionally relevant structural changes in PilM.

PubMed: 27022027
DOI: 10.1074/jbc.M116.718353

Experimental method

X-RAY DIFFRACTION (2.4 Å)