PKA from <i>Saccharomyces cerevisiae</i> can be activated by cyclic AMP and cyclic GMP

Cytryńska, Małgorzata; Wojda, Iwona; Frajnt, Magdalena; Jakubowicz, Teresa

doi:10.1139/w98-214

Cited by 18 publications

(14 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We have shown that phosphorylation of the 58-kDa protein by PKA can also be stimulated by cGMP. These results con¢rmed our earlier observations that native PKA from yeast can be stimulated by cAMP and cGMP [18].…”

Section: Resultssupporting

confidence: 92%

“…Postribosomal cell-free extracts were prepared in bu¡er A containing: 50 mM Tris^HCl pH 7.5, 6 mM 2-mercaptoethanol, 10 mM L-glycerophosphate, 1 mM phenylmethylsulfonyl £uoride (PMSF), 0.5 mM ethylenediaminetetraacetic acid (EDTA) as described previously [18]. Crude PKA preparation was obtained from cell-free extract by ammonium sulfate precipitation at 55% saturation.…”

Section: Puri¢cation Of Yeast Pkamentioning

confidence: 99%

“…PKA-2 preparation obtained after DEAE-cellulose was puri¢ed in parallel, using P-cellulose, hydroxylapatite and gel ¢ltration chromatography as described previously [18]. It is worth mentioning here that a protein of 58 kDa copuri¢ed with the PKA-1 fraction on all puri¢cation procedures used.…”

Section: Puri¢cation Of Yeast Pkamentioning

confidence: 99%

See 2 more Smart Citations

Saccharomyces cerevisiae pyruvate kinase Pyk1 is PKA phosphorylation substrate in vitro

Cytrynska¹

2001

FEMS Microbiology Letters

View full text Add to dashboard Cite

Fractionation of Saccharomyces cerevisiae postribosomal extract on DEAE-cellulose revealed two fractions of cAMP-dependent protein kinase (PKA-1 and PKA-2). The presence of PKA in both fractions was confirmed by immunoblotting with anti-Bcy1 antibodies. Yeast pyruvate kinase Pyk1 identified by amino acid microsequencing analysis and immunoblotting with anti-Pyk1 antibodies copurified with the PKA-1 but not the -2 fraction. Pyk1 can be phosphorylated by yeast PKA in vitro in the presence of cAMP and cGMP. Two-dimensional gel electrophoretic analysis revealed four phosphorylated forms of Pyk1 modified by PKA. In phosphorylation of Pyk1 mainly the Tpk2 catalytic subunit of yeast PKA was involved. ß

show abstract

Section: Resultssupporting

confidence: 92%

Section: Puri¢cation Of Yeast Pkamentioning

confidence: 99%

See 1 more Smart Citation

Saccharomyces cerevisiae pyruvate kinase Pyk1 is PKA phosphorylation substrate in vitro

Cytrynska¹

2001

FEMS Microbiology Letters

View full text Add to dashboard Cite

show abstract

“…7). Experimental evidence indicates that the subunit from Saccharomyces can be activated by both cAMP and cGMP (Cytrynska et al 1999). …”

Section: Familymentioning

confidence: 99%

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

“…The second messenger cAMP is well known to activate PKA by binding to regulatory subunits. Although, little is known regarding cGMP binding to bovine PKA, PKA is activated by both cAMP and cGMP in Orconectus limosis, Amblyomma americanum, Saccharomyces cerevisiae, and Pichia pastoris [20][21][22][23]. We find that both cAMP and cGMP bind to bovine PKA, with a preference for cAMP.…”

Section: Application Of Cofactor Fingerprinting With Std Nmr Inmentioning

confidence: 81%

Cofactor fingerprinting with STD NMR to characterize proteins of unknown function: identification of a rare cCMP cofactor preference

Yao

Sem

2004

FEBS Letters

View full text Add to dashboard Cite

Proteomics efforts have created a need for better strategies to functionally categorize newly discovered proteins. To this end, we have employed saturation transfer difference NMR with pools of closely related cofactors, to determine cofactor preferences. This approach works well for dehydrogenases and has also been applied to cyclic nucleotide-binding proteins. In the latter application, a protein (radial spoke protein-2, RSP2) that plays a central role in forming the radial spoke of Chlamydomonas reinhardtii flagella was shown to bind cCMP. cCMP-binding proteins are rare, although previous reports of their presence in sperm and flagella suggest that cCMP may have a more general role in flagellar function. 31 P NMR was used to monitor the preferential hydrolysis of ATP versus GTP, suggesting that RSP2 is a kinase.

show abstract

PKA from Saccharomyces cerevisiae can be activated by cyclic AMP and cyclic GMP

Cited by 18 publications

References 27 publications

Saccharomyces cerevisiae pyruvate kinase Pyk1 is PKA phosphorylation substrate in vitro

Saccharomyces cerevisiae pyruvate kinase Pyk1 is PKA phosphorylation substrate in vitro

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

Cofactor fingerprinting with STD NMR to characterize proteins of unknown function: identification of a rare cCMP cofactor preference

Contact Info

Product

Resources

About