cGMP-binding Prepares PKG for Substrate Binding by Disclosing the C-terminal Domain

Alverdi, Vera; Mazon, Hortense; Versluis, Cees; Hemrika, Wieger; Esposito, Gennaro; Heuvel, Robert van den; Scholten, Arjen; Heck, Albert J. R.

doi:10.1016/j.jmb.2007.11.053

Cited by 73 publications

(94 citation statements)

References 73 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One potentially confusing finding is that Rp-8AET-cGMPS, which inhibits PKGI kinase activity by preventing disengagement of the kinase and autoinhibitory sites, also disrupts normal LZ-dependent PKGI␣-RhoA binding. Others have shown, however, that liberation of the PKI␣ kinase domain from the autoinhibitory site leads to increased solubility of the LZ domain (25). Therefore, we interpret our findings to support that cGMP-mediated positioning of the PKGI␣ LZ domain, as well as its kinase activating effect, mediates PKGI␣ RhoA binding.…”

Section: Discussionsupporting

confidence: 83%

Direct Binding and Regulation of RhoA Protein by Cyclic GMP-dependent Protein Kinase Iα

Kato¹,

Blanton²,

Guang-rong

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Regulation of vascular smooth muscle tone by cGMP-dependent protein kinase I␣ (PKGI␣) through RhoA inhibition is not understood. Results: PKGI␣ binds RhoA, through cGMP-induced binding, and inhibition of RhoA requires PKGI␣ activity. Conclusion: RhoA-PKGI␣ co-interaction regulates RhoA function. Significance: Understanding RhoA regulation by PKGI␣ is important for understanding the regulation of vascular tone in vivo and in human diseases such as hypertension.

show abstract

Section: Discussionsupporting

confidence: 83%

Direct Binding and Regulation of RhoA Protein by Cyclic GMP-dependent Protein Kinase Iα

Kato¹,

Blanton²,

Guang-rong

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…1a), and it has been previously identified as a critical control unit for auto-inhibition and cGMP selectivity in PKG I␤ (17,18). In addition, we examined the CNB-B domain because although the CNB-A domain contributes to the cyclic nucleotide modulation of PKG I function (17,(35)(36)(37), the CNB-A domain undergoes only minor structural changes upon cyclic nucleotide binding (2,17). We found that the partial agonism of cAMP cannot be explained by a simple reversal of the two-state inactive/active conformational equilibrium that rationalizes cGMP activation (2,17).…”

mentioning

confidence: 99%

Mechanism of cAMP Partial Agonism in Protein Kinase G (PKG)

VanSchouwen¹,

Selvaratnam²,

Giri³

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Protein kinase G (PKG) is selectively activated by cGMP, but the mechanism of cGMP-versus-cAMP selectivity is not fully understood. Results: cAMP-bound PKG exists in a dynamic exchange between three states, inactive, active, and a partially autoinhibited state, which results in partial agonism. Conclusion: Partial agonism contributes to cGMP-versus-cAMP selectivity. Significance: The cGMP-versus-cAMP selectivity controls the cross-talk between cGMP-and cAMP-dependent signaling pathways.

show abstract

“…In one example of this, the application of IMS to the 11-membered TRAP complex, which was shown to retain its ring topology after the electrospray process, and that the binding of tryptophan aided in the preservation of this complex [37]. A more recent application of a traveling wave IMS instrument was to the analysis of active and inactive forms of the (cGMP)-dependant protein kinase by the Heck group [38]. Results obtained from this experiment suggest that the active cGMP bound protein adopts larger cross-sections than the inactive apo-protein, consistent with the expected opening of the structure implied by the HDX experiments conducted in the same study.…”

mentioning

confidence: 99%

Collision induced unfolding of protein ions in the gas phase studied by ion mobility-mass spectrometry: The effect of ligand binding on conformational stability

Hopper

Oldham

2009

J. Am. Soc. Mass Spectrom.

180

203

View full text Add to dashboard Cite

Ion mobility spectrometry, with subsequent mass spectrometric detection, has been employed to study the stability of compact protein conformations of FK-binding protein, hen egg-white lysozyme, and horse heart myoglobin in the presence and absence of bound ligands. Protein ions, generated by electrospray ionization from ammonium acetate buffer, were activated by collision with argon gas to induce unfolding of their compact structures. The collisional cross sections (⍀) of folded and unfolded conformations were measured in the T-Wave mobility cell of a Waters Synapt HDMS (Waters, Altrincham, UK) employing a calibration against literature values for a range of protein standards. In the absence of activation, collisional cross section measurements were found to be consistent with those predicted for folded protein structures. Under conditions of defined collisional activation energies partially unfolded conformations were produced. The degree of unfolding and dissociation induced by these defined collision energies are related to the stability of noncovalent intra-and intermolecular interactions within protein complexes. These findings highlight the additional conformational stability of protein ions in the gas phase resulting from ligand binding. E lectrospray ionization-mass spectrometry (ESI-MS) is a technique able to preserve the non-covalent interactions of protein-ligand complexes in the gas phase [1][2][3]. Since its original discovery, the application of ESI-MS in this area has accelerated rapidly [4,5]. The ESI-MS approach can provide a sensitive and efficient means of obtaining valuable information relevant to binding events allowing, for example, the stoichiometries of noncovalent complexes to be easily obtained [6,7]. The practical information available from ESI-MS measurement is already well documented [8]. In addition to stoichiometry, the affinities of protein-ligand interactions can be quantified using MS [9 -12]. In many cases, binding affinities determined using ESI-MS show good agreement with values obtained using other means, potentially validating MS methods for use in early-stage screening in the drug discovery process [13,14]. ESI-MS is not only suitable for the analysis of protein-ligand interactions, but has widespread utility in studying large protein-protein complexes [15,16], and has been applied to the preservation and detection of very large biomolecular assemblies, including the ribosome [17,18] and the tobacco mosaic virus (Ͼ40 MDa) [19].A key underlying issue in ESI-MS studies of ligand binding is to what extent these findings can be related to solution behavior. Given the great importance of water to the protein fold [20], it seems clear that desolvation should result in catastrophic loss of native structure, with accompanying consequences for ligand binding. Not withstanding the role of solvation spheres around the protein, a recent study has provided evidence that an interior water molecule in FK-binding protein makes significant contribution to the structural integrity of the fo...

show abstract

cGMP-binding Prepares PKG for Substrate Binding by Disclosing the C-terminal Domain

Cited by 73 publications

References 73 publications

Direct Binding and Regulation of RhoA Protein by Cyclic GMP-dependent Protein Kinase Iα

Direct Binding and Regulation of RhoA Protein by Cyclic GMP-dependent Protein Kinase Iα

Mechanism of cAMP Partial Agonism in Protein Kinase G (PKG)

Collision induced unfolding of protein ions in the gas phase studied by ion mobility-mass spectrometry: The effect of ligand binding on conformational stability

Contact Info

Product

Resources

About