9G1W

NMR solution structure of the Thermus thermophilus PilF-GSPIIA domain

Summary for 9G1W

• Entry DOI: 10.2210/pdb9g1w/pdb
• NMR Information: BMRB: 34929
• Descriptor: Type IV pilus assembly ATPase PilB (1 entity in total)
• Functional Keywords: pilt-class, gspii, pilf, motor protein
• Biological source: Thermus thermophilus HB27
• Total number of polymer chains: 1
• Total formula weight: 16924.60

Authors

Neissner, K.,Woehnert, J.,Hacker, C. (deposition date: 2024-07-10, release date: 2025-05-21)

Primary citation

Neissner, K.,Frohnapfel, C.,Keller, H.,Duchardt-Ferner, E.,Schneider, V.,Kamjou, Z.,Averhoff, B.,Wohnert, J.
NMR Solution Structure of the N-Terminal GSPII Domain from the Thermus Thermophilus Traffic ATPase PilF and Reconstruction of its c-di-GMP Binding Capability.
Chembiochem, 26:e202400959-e202400959, 2025

PubMed Abstract: The cyclic dinucleotide c-di-GMP is an important second messenger molecule in bacteria and interacts with a variety of receptor molecules including RNA and protein domains. An important class of c-di-GMP-binding protein domains are the general secretory pathway type II (GSPII) domains as exemplified by the N-terminal domain of the ATPase MshE from Vibrio cholerae (MshEN). MshEN binds monomeric c-di-GMP via two consecutive copies of a 24-residue sequence motif, which form a compact 4-α-helical bundle. The ATPase PilF from Thermus thermophilus regulates pilus formation, motility and DNA-uptake. Its N-terminal section contains three consecutive GSPII domains (GSPII-A-GSPII-C) all with considerable sequence homology to MshEN. While the GSPII-B and the GSPII-C domains bind c-di-GMP, the GSPII-A domain does not. To determine why it is incapable of c-di-GMP-binding we determined the NMR-solution structure of this domain. Our structure shows how small deviations in the consensus motif sequence, a stabilizing N-terminal helical capping motif and intersubdomain interactions absent in MshEN cooperate to prevent c-di-GMP-binding. By combining point mutations and truncations, we re-established the c-di-GMP binding capability. Our findings shed new light on the evolution and functional diversification of GSPII domains and the importance of sequence variations for protein activity in this domain family.

PubMed: 39960869
DOI: 10.1002/cbic.202400959

Experimental method

SOLUTION NMR