Regulatory Subunit Of Protein Kinase A: Structure of Deletion Mutant with cAMP Binding Domains

Su, Yi-Hsien; Dostmann, Wolfgang R.; Herberg, Friedrich W.; Durick, Kyle; Xuong, N.-H.; Eyck, Lynn Ten; Taylor, Susan S.; Varughese, Kottayil I.

doi:10.1126/science.7638597

Cited by 370 publications

(505 citation statements)

References 53 publications

Supporting

Mentioning

486

Contrasting

Order By: Relevance

“…Priority was set to placing gaps within loops connecting secondary structure elements. The cAMP-binding regions were manually curated with the program GeneDoc (Nicholas et al 1997), taking into consideration the crystal structures of bovine RI␣ (Su et al 1995) and rat RII␤ (Diller et al 2001). The Gonnet weight matrix (Gonnet et al 1994) was used in the sequence alignments.…”

Section: Methodsmentioning

confidence: 99%

“…5). The alignment of the cAMP-binding domains was curated based on structural evidence from the crystal structures of bovine RI␣ (Su et al 1995) and rat RII␤ (Diller et al 2001). Most of the variability in the cAMP-binding domains corresponded to the loop between ␤-strand 4 and ␤-strand 5 of both domain A and domain B, and the C-terminal region.…”

Section: Sequence Alignment and Phylogenetic Analysismentioning

confidence: 99%

“…The PBCs are defined as the segments of each cAMPbinding domain that contain most of the key residues for cAMP-mediated activation of PKA (Figs. 1B and C and 5) (Su et al 1995;Diller et al 2001).…”

Section: Sequence Alignment and Phylogenetic Analysismentioning

confidence: 99%

See 2 more Smart Citations

Classification and Phylogenetic Analysis of the cAMP-Dependent Protein Kinase Regulatory Subunit Family

2002

View full text Add to dashboard Cite

Abstract. The members of the PKA regulatory subunit family (PKA-R family) were analyzed by multiple sequence alignment and clustering based on phylogenetic tree construction. According to the phylogenetic trees generated from multiple sequence alignment of the complete sequences, the PKA-R family was divided into four subfamilies (types I to IV). Members of each subfamily were exclusively from animals (types I and II), fungi (type III), and alveolates (type IV). Application of the same methodology to the cAMP-binding domains, and subsequently to the region delimited by ␤-strands 6 and 7 of the crystal structures of bovine RI␣ and rat RII␤ (the phosphate-binding cassette; PBC), proved that this highly conserved region was enough to classify unequivocally the members of the PKA-R family. A single signature sequence, F-G-E-[LIV]-A-L-[LIMV]-x(3)-[PV]-R-[ANQV]-A, corresponding to the PBC was identified which is characteristic of the PKA-R family and is sufficient to distinguish it from other members of the cyclic nucleotide-binding protein superfamily. Specific determinants for the A and B domains of each Rsubunit type were also identified. Conserved residues defining the signature motif are important for interaction with cAMP or for positioning the residues that directly interact with cAMP. Conversely, residues that define subfamilies or domain types are not conserved and are mostly located on the loop that connects ␣-helix BЈ and ␤ strand 7.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Sequence Alignment and Phylogenetic Analysismentioning

confidence: 99%

See 1 more Smart Citation

Classification and Phylogenetic Analysis of the cAMP-Dependent Protein Kinase Regulatory Subunit Family

2002

View full text Add to dashboard Cite

show abstract

“…When four molecules of cAMP bind the regulatory subunit dimer (two to each subunit) there is a conformational change in PKA which results in lower affinity for the catalytic subunit and the complex dissociates. The regulatory subunit possesses two cAMP binding sites (known as "A" and "B") that act cooperatively (Su et al, 1995). It is not clear which isoforms of PKA are present in human β cells: however both PKA type I and II have been isolated from DEAE-cellulose ion-exchange chromatography of rat islets (Sugden et al, 1979).…”

Section: Activation Of Pkamentioning

confidence: 99%

Mechanisms of action of glucagon-like peptide 1 in the pancreas

Doyle

Egan

2007

Pharmacology & Therapeutics

561

465

View full text Add to dashboard Cite

Glucagon-like peptide-1 is a hormone that is encoded in the proglucagon gene. It is mainly produced in enteroendocrine L cells of the gut and is secreted into the blood stream when food containing fat, protein hydrolysate and/or glucose enters the duodenum. Its particular effects on insulin and glucagon secretion have generated a flurry of research activity over the past twenty years culminating in a naturally occurring GLP-1 receptor agonist, exendin-4, now being used to treat type 2 diabetes. GLP-1 engages a specific G-protein coupled receptor that is present in tissues other than the pancreas (brain, kidney, lung, heart, major blood vessels). The most widely studied cell activated by GLP-1 is the insulin-secreting beta cell where its defining action is augmentation of glucose-induced insulin secretion. Upon GLP-1 receptor activation, adenylyl cyclase is activated and cAMP generated, leading, in turn, to cAMP-dependent activation of second messenger pathways, such as the PKA and Epac pathways. As well as short-term effects of enhancing glucose-induced insulin secretion, continuous GLP-1 receptor activation also increases insulin synthesis, and beta cell proliferation and neogenesis. Although these latter effects cannot be currently monitored in humans, there are substantial improvements in glucose tolerance and increases in both first phase and plateau phase insulin secretory responses in type 2 diabetic patients treated with exendin-4. This review we will focus on the effects resulting from GLP-1 receptor activation in islets of Langerhans

show abstract

“…The C199A mutant and wildtype C-subunit showed identical binding profiles with FAM-IP20 (data not shown), while the binding and activation behavior of the deletion mutant is nearly identical to wildtype RIα 27 The C199A mutant of recombinant murine catalytic subunit 28 and bovine RIα(Δ1-91) 29 were expressed and purified from E. coli as previously described. …”

Section: Protein Expression and Purificationmentioning

confidence: 98%

Assay Principle for Modulators of Protein−Protein Interactions and Its Application to Non-ATP-Competitive Ligands Targeting Protein Kinase A

et al. 2006

View full text Add to dashboard Cite

Targeting sites that modulate protein-protein interactions represents an ongoing challenge for drug discovery. We have devised an assay principle, named Ligand-Regulated Competition (LiReC), in an effort to find non-ATP competitive small molecule regulators for type Iα cAMP-dependent Protein Kinase A (PKA-Iα), a protein complex that is implicated in disease. Our assay based on the LiReC principle utilizes a competitive fluorescent peptide probe to assess the integrity of the PKA-Iα complex upon introduction of an allosteric ligand. The developed fluorescence polarization method screens for small molecules that specifically protect (antagonists) or conversely activate (agonists) this protein complex. In high throughput format, various cyclic nucleotide-derived agonists and antagonists are successfully detected with high precision. Furthermore, assay performance (Z'-factors above 0.7) far exceeds the minimum requirement for small molecule screening. To identify compounds that operate through novel modes of action, our method shields the ATP binding site and purposely excludes ATP-competitive ligands. These proof-of-principle experiments highlight the potential of the LiReC technique and suggest its application to other protein complexes, thereby providing a novel approach to identify and characterize modulators (small molecules, proteins, peptides, or nucleic acids) of protein-protein systems.

show abstract

Regulatory Subunit Of Protein Kinase A: Structure of Deletion Mutant with cAMP Binding Domains

Cited by 370 publications

References 53 publications

Classification and Phylogenetic Analysis of the cAMP-Dependent Protein Kinase Regulatory Subunit Family

Classification and Phylogenetic Analysis of the cAMP-Dependent Protein Kinase Regulatory Subunit Family

Mechanisms of action of glucagon-like peptide 1 in the pancreas

Assay Principle for Modulators of Protein−Protein Interactions and Its Application to Non-ATP-Competitive Ligands Targeting Protein Kinase A

Contact Info

Product

Resources

About