Expression, purification, and characterization of the cGMP‐dependent protein kinases Iβ and II using the baculovirus system

  title={Expression, purification, and characterization of the cGMP‐dependent protein kinases I$\beta$ and II using the baculovirus system},
  author={D P{\"o}hler and Elke Butt and Joseph Meissner and S M{\"u}ller and Martin J. Lohse and Ulrich Walter and Suzanne M. Lohmann and Thomas Jarchau},
  journal={FEBS Letters},

Endogenous Type II cGMP-dependent Protein Kinase Exists as a Dimer in Membranes and Can Be Functionally Distinguished from the Type I Isoforms*

Ex vivo endogenous cGK II displays a distinct activation/inhibition profile with respect to cGMP analogs, which permits a pharmacological dissection between cGk II- andcGK I-mediated physiological processes.

Diversity of cAMP-dependent protein kinase isoforms and their anchoring proteins in mouse ventricular tissue.

A proposed fusion protein of paralemmin 2 and AKAP2 could be decisively established as a novel AKAP at the protein level in ventricular tissue and the improved chemical proteomics screen offers a broad, but detailed, view on nature's complex diversity in cyclic nucleotide signaling mechanisms.

cGMP-dependent Protein Kinase Iβ Physically and Functionally Interacts with the Transcriptional Regulator TFII-I*

TFII-I by itself had little effect on a full-length fospromoter in baby hamster kidney cells, but it synergistically enhanced transcriptional activation by G-kinase Iβ, an unusual transcriptional regulator that associates with multiple proteins to modulate both basal and signal-induced transcription.

cGMP-Dependent Protein Kinase Inhibitors in Health and Disease

It will be discussed that the distinct inhibition of the PKG enzymes could be used as a valuable pharmacological target e.g., in the treatment of cardiovascular diseases, diarrhea, cancer or malaria.

Interaction of cCMP with the cGK, cAK and MAPK kinases in murine tissues

This work investigated the effect of cCMP on purified cyclic nucleotide-dependent protein kinases and lung and jejunum tissues of wild type (WT), cGKI-knockout (cGKI KO) and cGKII-knocking out (cKII KO) mice and found no stimulation of phosphorylation in KO tissue lysates.

Actin Binding of Human LIM and SH3 Protein Is Regulated by cGMP- and cAMP-dependent Protein Kinase Phosphorylation on Serine 146*

Data suggest that phosphorylation of LASP by cGK and cAK may be involved in cytoskeletal organization and cell motility.

Selectivity in Enrichment of cAMP-dependent Protein Kinase Regulatory Subunits Type I and Type II and Their Interactors Using Modified cAMP Affinity Resins*S

A combination of affinity chromatography and mass spectrometry-based quantitative proteomics is employed to investigate specificity in PKA-AKAP interactions and provides the first insights into their specificity.

Functional analysis of cGMP-dependent protein kinases I and II as mediators of NO/cGMP effects

The analysis of vasodilator-stimulated phosphoprotein (VASP) phosphorylation by polyclonal antibodies and newly developed monoclonal antibodies allows the quantitative measurement of cGK activity in intact cells.



Properties of a cGMP-dependent monomeric protein kinase from bovine aorta.

The results indicated that the catalytic domain, cGMP binding domain(s), and inhibitory domain of cGK interact primarily within the same subunit rather than between subunits of the dimer as previously hypothesized for dimeric c GK.

The activation of expressed cGMP‐dependent protein kinase isozymes Iα and Iβ is determined by the different amino‐termini

It is suggested that a change from the I alpha amino-terminal domain to that of I beta increases the apparent KA value for cGMP 10-fold by altering the binding properties of binding site 1.

Detection of cGMP dependent protein kinase isozymes by specific antibodies.

Polyclonal antibodies were developed which recognize both isozymes or specifically the I alpha and I beta isoform of cGMP-dependent protein kinase, and yielded strong signals with extracts of trachea, aorta, stomach and uterus.

[1] Assays of protein kinase

Endogenous expression of type II cGMP-dependent protein kinase mRNA and protein in rat intestine. Implications for cystic fibrosis transmembrane conductance regulator.

Viewed collectively, the data suggest that cGK II is the mediator of STa and cGMP effects on Cl- transport in intestinal-epithelia.

Immunological distinction between guanosine 3':5'-monophosphate-dependent and adenosine 3':5'-monophosphate-dependent protein kinases.

The substantial differences between the immunological properties of the cGMP-dependent and cAMP-dependent protein kinases suggest that these two enzymes have distinct physiological roles.