5K94

Deletion-Insertion Chimera of MBP with the Preprotein Cross-Linking Domain of the SecA ATPase

Summary for 5K94

• Entry DOI: 10.2210/pdb5k94/pdb
• Descriptor: Maltose-binding periplasmic protein,Protein translocase subunit SecA,Maltose-binding periplasmic protein, 2-[3-(2-HYDROXY-1,1-DIHYDROXYMETHYL-ETHYLAMINO)-PROPYLAMINO]-2-HYDROXYMETHYL-PROPANE-1,3-DIOL (3 entities in total)
• Functional Keywords: preprotein translocase, seca, preprotein cross-linking domain, ppxd, mbp chimera, protein transport
• Biological source: Escherichia coli O157:H7; More
• Total number of polymer chains: 2
• Total formula weight: 113418.00

Authors

Shilton, B.H.,Hackett, J.,Ghonaim, N. (deposition date: 2016-05-31, release date: 2017-06-07, Last modification date: 2023-09-27)

Primary citation

Khalili Yazdi, A.,Namjoshi, S.,Hackett, J.,Ghonaim, N.,Shilton, B.H.
Characterization of a polypeptide-binding site in the DEAD Motor of the SecA ATPase.
FEBS Lett., 591:3378-3390, 2017

PubMed Abstract: We coupled peptides from a CNBr digest of signal-sequenceless maltose-binding protein (MBP) to a surface plasmon resonance chip. SecA-N95, SecA-N68, and SecA-DM (which consists of only the DEAD Motor domains NBD1 and NBD2) bound to the immobilized peptides; ADP weakened the binding. SecA-DM, which lacks the 'preprotein cross-linking domain' (PPXD), displayed the most extensive binding, while an MBP-PPXD chimera showed no binding, demonstrating that the PPXD does not contribute to the binding. We characterized the sequence specificity using oriented peptide libraries; these results enabled synthesis of a 20-residue peptide that was used to recapitulate the results obtained with MBP-derived peptides. This study shows that there is a promiscuous and nucleotide-modulated peptide-binding site in the DEAD Motor domains of SecA.

PubMed: 28862749
DOI: 10.1002/1873-3468.12832

Experimental method

X-RAY DIFFRACTION (2.1 Å)