Scanning calorimetric study of the thermal unfolding of catabolite activator protein from Escherichia coli in the absence and presence of cyclic mononucleotides.


Abstract

The thermal unfolding of the catabolite activator protein (CAP) of Escherichia coli and the complexes it forms with adenosine cyclic 3',5'-phosphate (cAMP) and guanosine cyclic 3',5'-phosphate (cGMP) was studied by high-sensitivity differential scanning calorimetry (DSC). The thermal denaturation of CAP at pH 7.00 gave an irreversible, symmetrical denaturation curve with a single peak. Distinctly different, more complex DSC curves were obtained for the thermal denaturation of the cAMP-protein and cGMP-protein complexes. The DSC data indicate intermolecular cooperation among CAP dimers, with the extent of oligomerization remaining unchanged during unfolding of the protein. The DSC curves for the thermal denaturation of the cAMP-protein complex and cGMP-protein complex have been resolved into three and two components, respectively, according to the model of independent two-state processes. Analysis of the DSC data suggests two and three independent domains for cGMP-protein and cAMP-protein complexes, respectively, with dissociation of mononucleotide occurring in the second component in both cases during protein denaturation. Furthermore, our studies indicate that the presence of either ligand alters the degree of oligomerization of CAP dimers, cAMP having a greater effect than cGMP. Study holds ProTherm entries: 3845 Extra Details: additive : EDTA(0.2 mM), intermolecular cooperation; oligomerization;,three independent domains; protein denaturation

Submission Details

ID: MTJtwmvu3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:23 p.m.

Version: 1

Publication Details
Ghosaini LR;Brown AM;Sturtevant JM,Biochemistry (1988) Scanning calorimetric study of the thermal unfolding of catabolite activator protein from Escherichia coli in the absence and presence of cyclic mononucleotides. PMID:2844254
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
4BHP 2013-05-08 A structural model of CAP mutant (T127L and S128I) in cGMP-bound state
4BH9 2013-05-08 A structural model of CAP mutant (T127L and S128I) in the apo state
2WC2 2009-04-21 Nmr structure of catabolite activator protein in the unliganded state
4R8H 2015-07-22 1.46 The role of protein-ligand contacts in allosteric regulation of the Escherichia coli Catabolite Activator Protein
4HZF 2013-10-30 1.48 structure of the wild type Catabolite gene Activator Protein
4I0B 2013-10-30 1.5 structure of the mutant Catabolite gene activator protein H160L
3RYP 2012-05-02 1.6 Domain-domain flexibility leads to allostery within the camp receptor protein (CRP)
3KCC 2009-11-17 1.66 Crystal structure of D138L mutant of Catabolite Gene Activator Protein
4I02 2013-10-30 1.75 structure of the mutant Catabolite gene activator protein V140A
1HW5 2001-01-17 1.82 THE CAP/CRP VARIANT T127L/S128A
1I5Z 2003-06-17 1.9 STRUCTURE OF CRP-CAMP AT 1.9 A
3QOP 2012-04-04 1.96 Domain-domain flexibility leads to allostery within the camp receptor protein (CRP)
4FT8 2013-12-18 1.97 E. coli Catabolite Activator Protein with Cobalt and Sulfate Ligands
3IYD 2009-11-10 19.8 Three-dimensional EM structure of an intact activator-dependent transcription initiation complex
4I09 2013-10-30 2.05 structure of the mutant Catabolite gene activator protein V132L
3ROU 2012-05-02 2.1 Domain-domain flexibility leads to allostery within the camp receptor protein (CRP)
1ZRF 2006-03-21 2.1 4 crystal structures of CAP-DNA with all base-pair substitutions at position 6, CAP-[6C;17G]ICAP38 DNA
1G6N 2000-12-15 2.1 2.1 ANGSTROM STRUCTURE OF CAP-CAMP
4I0A 2013-10-30 2.2 structure of the mutant Catabolite gene activator protein V132A
1I6X 2003-06-17 2.2 STRUCTURE OF A STAR MUTANT CRP-CAMP AT 2.2 A
2CGP 1998-02-04 2.2 CATABOLITE GENE ACTIVATOR PROTEIN/DNA COMPLEX, ADENOSINE-3',5'-CYCLIC-MONOPHOSPHATE
2GZW 2007-05-15 2.21 Crystal structure of the E.coli CRP-cAMP complex
4I01 2013-10-30 2.3 Structure of the mutant Catabolite gen activator protein V140L
3FWE 2009-09-08 2.3 Crystal Structure of the Apo D138L CAP mutant
1J59 2002-03-01 2.5 CATABOLITE GENE ACTIVATOR PROTEIN (CAP)/DNA COMPLEX + ADENOSINE-3',5'-CYCLIC-MONOPHOSPHATE
3RPQ 2012-05-02 2.61 Domain-domain flexibility leads to allostery within the camp receptor protein (CRP)
1RUN 1997-01-10 2.7 CATABOLITE GENE ACTIVATOR PROTEIN (CAP)/DNA COMPLEX + ADENOSINE-3',5'-CYCLIC-MONOPHOSPHATE
1RUO 1997-01-10 2.7 CATABOLITE GENE ACTIVATOR PROTEIN (CAP) MUTANT/DNA COMPLEX + ADENOSINE-3',5'-CYCLIC-MONOPHOSPHATE
3RYR 2012-05-02 2.7 Domain-domain flexibility leads to allostery within the camp receptor protein (CRP)
1ZRE 2006-03-21 2.8 4 crystal structures of CAP-DNA with all base-pair substitutions at position 6, CAP-[6G;17C]ICAP38 DNA
1ZRD 2006-03-21 2.8 4 crystal structures of CAP-DNA with all base-pair substitutions at position 6, CAP-[6A;17T]ICAP38 DNA
1ZRC 2006-03-21 2.8 4 Crystal structures of CAP-DNA with all base-pair substitutions at position 6, CAP-ICAP38 DNA
1O3T 2003-04-08 2.8 PROTEIN-DNA RECOGNITION AND DNA DEFORMATION REVEALED IN CRYSTAL STRUCTURES OF CAP-DNA COMPLEXES
3RDI 2012-05-02 2.95 Domain-domain flexibility leads to allostery within the camp receptor protein (CRP)
3N4M 2011-05-25 2.99 E. coli RNA polymerase alpha subunit C-terminal domain in complex with CAP and DNA
1O3R 2003-04-08 3.0 PROTEIN-DNA RECOGNITION AND DNA DEFORMATION REVEALED IN CRYSTAL STRUCTURES OF CAP-DNA COMPLEXES
1O3S 2003-04-08 3.0 PROTEIN-DNA RECOGNITION AND DNA DEFORMATION REVEALED IN CRYSTAL STRUCTURES OF CAP-DNA COMPLEXES
1O3Q 2003-04-08 3.0 PROTEIN-DNA RECOGNITION AND DNA DEFORMATION REVEALED IN CRYSTAL STRUCTURES OF CAP-DNA COMPLEXES
1CGP 1994-01-31 3.0 CATABOLITE GENE ACTIVATOR PROTEIN (CAP)/DNA COMPLEX + ADENOSINE-3',5'-CYCLIC-MONOPHOSPHATE
1LB2 2002-09-06 3.1 Structure of the E. coli alpha C-terminal domain of RNA polymerase in complex with CAP and DNA
3HIF 2009-09-08 3.59 The crystal structure of apo wild type CAP at 3.6 A resolution.
6B6H 2017-11-15 3.9 The cryo-EM structure of a bacterial class I transcription activation complex
5CIZ 2016-06-29 5.01 E. coli RNA polymerase alpha subunit CTD in complex with CAP and DNA: A(5)-tract binding site for alpha CTD

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
99.5 cAMP-activated global transcriptional regulator CRP P0A2T6 CRP_SALTY
99.5 cAMP-activated global transcriptional regulator CRP P0A2T7 CRP_KLEAE
100.0 cAMP-activated global transcriptional regulator CRP P0ACK1 CRP_SHIFL
100.0 cAMP-activated global transcriptional regulator CRP P0ACJ8 CRP_ECOLI
100.0 cAMP-activated global transcriptional regulator CRP P0ACJ9 CRP_ECOL6
100.0 cAMP-activated global transcriptional regulator CRP P0ACK0 CRP_ECO57