Investigation into the interactions between diadenosine 5',5'''-P1,P4-tetraphosphate and two proteins: molecular chaperone GroEL and cAMP receptor protein.
Diadenosine 5',5'''-P(1),P(4)-tetraphosphate (Ap(4)A) is a dinucleoside polyphosphate found ubiquitously in eukaryotic and prokaryotic cells. Despite Ap(4)A being universal, its functions have proved to be difficult to define, although they appear to have a strong presence during cellular stress. Here we report on our investigations into the nature and properties of putative Ap(4)A interactions with Escherichia coli molecular chaperone GroEL and cAMP receptor protein (CRP). We confirm previous literature observations that GroEL is an Ap(4)A binding protein and go on to prove that binding of Ap(4)A to GroEL involves a set of binding sites (one per monomer) distinct from the well-known GroEL ATP/ADP sites. Binding of Ap(4)A to GroEL appears to enhance ATPase rates at higher temperatures, encourages the release of bound ADP, and may promote substrate protein release through differential destabilization of the substrate protein-GroEL complex. We suggest that such effects should result in enhanced GroEL/GroES chaperoning activities that could be a primary reason for the improved yields of the refolded substrate protein observed during GroEL/GroES-assisted folding and refolding at >or=30 degrees C in the presence of Ap(4)A. In contrast, we were unable to obtain any data to support a direct role for Ap(4)A interactions with CRP.
Tanner
Julian A
JA
Imperial College Genetic Therapies Centre, Department of Chemistry, Flowers Building, Armstrong Road, Imperial College London, UK.
Wright
Michael
M
Christie
E Margaret
EM
Preuss
Monika K
MK
Miller
Andrew D
AD
D001483:Base Sequence; D018834:Chaperonin 60 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002942:Circular Dichroism; D002373:Cyclic AMP Receptor Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015226:Dinucleoside Phosphates / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D011485:Protein Binding; D013696:Temperature
D018834:Chaperonin 60; D002373:Cyclic AMP Receptor Protein; D015226:Dinucleoside Phosphates; D018832:Molecular Chaperones; C020733:diadenosine tetraphosphate