The interaction between the chaperone SecB and its ligands: evidence for multiple subsites for binding.
The chaperone protein SecB is dedicated to the facilitation of export of proteins from the cytoplasm to the periplasm and outer membrane of Escherichia coli. It functions to bind and deliver precursors of exported proteins to the membrane-associated translocation apparatus before the precursors fold into their native stable structures. The binding to SecB is characterized by a high selectivity for ligands having nonnative structure but a low specificity for consensus in sequence among the ligands. A model previously presented (Randall LL, Hardy SJS, 1995, Trends Biochem Sci 20:65-69) to rationalize the ability of SecB to distinguish between the native and nonnative states of a polypeptide proposes that the SecB tetramer contains two types of subsites for ligand binding: one kind that would interact with extended flexible stretches of polypeptides and the other with hydrophobic regions. Here we have used titration calorimetry, analytical ultracentrifugation, and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry to obtain evidence that such distinguishable subsites exist.
Randall
L L
LL
Department of Biochemistry and Biophysics, Washington State University, Pullman 99164-4660, USA.
Hardy
S J
SJ
Topping
T B
TB
Smith
V F
VF
Bruce
J E
JE
Smith
R D
RD
D007611:Aprotinin / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D002151:Calorimetry; D017357:Cyclotrons; D004926:Escherichia coli / Q000737:chemistry; D005583:Fourier Analysis; D046911:Macromolecular Substances; D013058:Mass Spectrometry / Q000379:methods; D018832:Molecular Chaperones; D010455:Peptides / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D013816:Thermodynamics; D014461:Ultracentrifugation
D001426:Bacterial Proteins; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D010455:Peptides; C057842:SecB protein, Bacteria; D007611:Aprotinin; D012260:Ribonucleases; C019760:Bacillus amyloliquefaciens ribonuclease