2VII

PspF1-275-Mg-AMP

Summary for 2VII

• Entry DOI: 10.2210/pdb2vii/pdb
• Related: 2BJV 2BJW 2C96 2C98 2C99 2C9C
• Descriptor: PSP OPERON TRANSCRIPTIONAL ACTIVATOR, ADENOSINE MONOPHOSPHATE, MAGNESIUM ION, ... (4 entities in total)
• Functional Keywords: aaa, pspf, activator, atp-binding, dna-binding, sigma54 activator, nucleotide-binding, transcription, gene regulation, enhancer binding protein, transcription activation, transcription regulation, two-component regulatory system
• Biological source: ESCHERICHIA COLI
• Cellular location: Cytoplasm (Potential): P37344
• Total number of polymer chains: 1
• Total formula weight: 29762.19

Authors

Joly, N.,Rappas, M.,Buck, M.,Zhang, X. (deposition date: 2007-12-04, release date: 2008-01-22, Last modification date: 2023-12-13)

Primary citation

Joly, N.,Rappas, M.,Buck, M.,Zhang, X.
Trapping of a Transcription Complex Using a New Nucleotide Analogue: AMP Aluminium Fluoride
J.Mol.Biol., 375:1206-, 2008

PubMed Abstract: Mechanochemical proteins rely on ATP hydrolysis to establish the different functional states required for their biological output. Studying the transient functional intermediate states these proteins adopt as they progress through the ATP hydrolysis cycle is key to understanding the molecular basis of their mechanism. Many of these intermediates have been successfully 'trapped' and functionally characterised using ATP analogues. Here, we present a new nucleotide analogue, AMP-AlF(x), which traps PspF, a bacterial enhancer binding protein, in a stable complex with the sigma(54)-RNA polymerase holoenzyme. The crystal structure of AMP-AlF(x)*PspF(1-275) provides new information on protein-nucleotide interactions and suggests that the beta and gamma phosphates are more important than the alpha phosphate in terms of sensing nucleotide bound states. In addition, functional data obtained with AMP-AlF(x) establish distinct roles for the conserved catalytic AAA(+) (ATPases associated with various cellular activities) residues, suggesting that AMP-AlF(x) is a powerful new tool to study AAA(+) protein family members and, more generally, Walker motif ATPases.

PubMed: 18082766
DOI: 10.1016/J.JMB.2007.11.050

Experimental method

X-RAY DIFFRACTION (2.85 Å)