SASDDS4: cGMP-dependent protein kinase 1: ∆53 PKG Iα (cGMP-dependent prote...

Basic information

• Entry: Database: SASBDB / ID: SASDDS4

Sample

cGMP-dependent protein kinase 1: ∆53 PKG Iα

• cGMP-dependent protein kinase 1 (protein), ∆53 PKG Iα, Bos taurus

Function / homology

Function:

cGMP effects / cGMP-dependent protein kinase / cGMP-dependent protein kinase activity / Rap1 signalling / negative regulation of vascular associated smooth muscle cell migration / collateral sprouting / negative regulation of platelet aggregation / relaxation of vascular associated smooth muscle / regulation of GTPase activity / cGMP-mediated signaling / dendrite development / negative regulation of vascular associated smooth muscle cell proliferation / calcium channel regulator activity / cGMP binding / forebrain development / neuron migration / protein phosphorylation / protein serine kinase activity / Golgi apparatus / ATP binding / identical protein binding / plasma membrane / cytoplasm / cytosol

Domain/homology:

cGMP-dependent protein kinase, N-terminal coiled-coil domain / Coiled-coil N-terminus of cGMP-dependent protein kinase / cGMP-dependent kinase / cGMP-dependent protein kinase, catalytic domain / Cyclic nucleotide-binding domain signature 2. / Cyclic nucleotide-binding domain signature 1. / Cyclic nucleotide-binding, conserved site / Cyclic nucleotide-monophosphate binding domain / Cyclic nucleotide-binding domain / cAMP/cGMP binding motif profile. / Cyclic nucleotide-binding domain / Cyclic nucleotide-binding domain superfamily / Extension to Ser/Thr-type protein kinases / AGC-kinase, C-terminal / AGC-kinase C-terminal domain profile. / RmlC-like jelly roll fold / Serine/threonine-protein kinase, active site / Serine/Threonine protein kinases active-site signature. / Protein kinase domain / Serine/Threonine protein kinases, catalytic domain / Protein kinase, ATP binding site / Protein kinases ATP-binding region signature. / Protein kinase domain profile. / Protein kinase domain / Protein kinase-like domain superfamily

Component:

cGMP-dependent protein kinase 1

• Biological species: Bos taurus (cattle)
• Citation: Journal: J Biol Chem / Year: 2018; Title: An N-terminally truncated form of cyclic GMP-dependent protein kinase Iα (PKG Iα) is monomeric and autoinhibited and provides a model for activation.; Authors: Thomas M Moon / Jessica L Sheehe / Praveena Nukareddy / Lydia W Nausch / Jessica Wohlfahrt / Dwight E Matthews / Donald K Blumenthal / Wolfgang R Dostmann / ; Abstract: The type I cGMP-dependent protein kinases (PKG I) serve essential physiological functions, including smooth muscle relaxation, cardiac remodeling, and platelet aggregation. These enzymes form homodimers through their N-terminal dimerization domains, a feature implicated in regulating their cooperative activation. Previous investigations into the activation mechanisms of PKG I isoforms have been largely influenced by structures of the cAMP-dependent protein kinase (PKA). Here, we examined PKG Iα activation by cGMP and cAMP by engineering a monomeric form that lacks N-terminal residues 1-53 (Δ53). We found that the construct exists as a monomer as assessed by whole-protein MS, size-exclusion chromatography, and small-angle X-ray scattering (SAXS). Reconstruction of the SAXS 3D envelope indicates that Δ53 has a similar shape to the heterodimeric RIα-C complex of PKA. Moreover, we found that the Δ53 construct is autoinhibited in its cGMP-free state and can bind to and be activated by cGMP in a manner similar to full-length PKG Iα as assessed by surface plasmon resonance (SPR) spectroscopy. However, we found that the Δ53 variant does not exhibit cooperative activation, and its cyclic nucleotide selectivity is diminished. These findings support a model in which, despite structural similarities, PKG Iα activation is distinct from that of PKA, and its cooperativity is driven by in interactions between protomers.

Contact author

• Thomas Moon (University of Arizona, Tucson, AZ, USA)

Models

• Model #1814: ; Type: dummy / Software: (5.0) / Radius of dummy atoms: 3.25 A / Symmetry: p1 / Comment: Filtered, Averaged 3-D envelope / Chi-square value: 0.602 / P-value: 0.999000; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: cGMP-dependent protein kinase 1: ∆53 PKG Iα
• Buffer: Name: 50 mM MES, 300 mM NaCl, 1 mM TCEP, 5 mM DTT / pH: 6.9

Entity #988

Name: ∆53 PKG Iα / Type: protein / Description: cGMP-dependent protein kinase 1 / Formula weight: 70.433 / Num. of mol.: 1 / Source: Bos taurus / References: UniProt: P00516
Sequence:

GAMDIGPRTT RAQGISAEPQ TYRSFHDLRQ AFRKFTKSER SKDLIKEAIL DNDFMKNLEL SQIQEIVDCM YPVEYGKDSC IIKEGDVGSL VYVMEDGKVE VTKEGVKLCT MGPGKVFGEL AILYNCTRTA TVKTLVNVKL WAIDRQCFQT IMMRTGLIKH TEYMEFLKSV PTFQSLPEEI LSKLADVLEE THYENGEYII RQGARGDTFF IISKGKVNVT REDSPNEDPV FLRTLGKGDW FGEKALQGED VRTANVIAAE AVTCLVIDRD SFKHLIGGLD DVSNKAYEDA EAKAKYEAEA AFFANLKLSD FNIIDTLGVG GFGRVELVQL KSEESKTFAM KILKKRHIVD TRQQEHIRSE KQIMQGAHSD FIVRLYRTFK DSKYLYMLME ACLGGELWTI LRDRGSFEDS TTRFYTACVV EAFAYLHSKG IIYRDLKPEN LILDHRGYAK LVDFGFAKKI GFGKKTWTFC GTPEYVAPEI ILNKGHDISA DYWSLGILMY ELLTGSPPFS GPDPMKTYNI ILRGIDMIEF PKKIAKNAAN LIKKLCRDNP SERLGNLKNG VKDIQKHKWF EGFNWEGLRK GTLTPPIIPS VASPTDTSNF DSFPEDNDEP PPDDNSGWDI DF

Experimental information

• Beam: Instrument name: Stanford Synchrotron Radiation Lightsource (SSRL) BL4-2; City: Stanford, CA / 国: USA / Type of source: X-ray synchrotron / Wavelength: 0.1127 Å / Dist. spec. to detc.: 1.7 mm
• Detector: Name: Rayonix MX225-HE

Scan

Title: cGMP-dependent protein kinase 1: ∆53 PKG Iα / Measurement date: Jun 6, 2015 / Storage temperature: 4 °C / Cell temperature: 22 °C / Exposure time: 1 sec. / Number of frames: 600 / Unit: 1/A /

	Min	Max
Q	0.0087	0.5136

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 192 /

	Min	Max
Q	0.0160014	0.190375
P(R) point	1	192
R	0	96.77

Result

Type of curve: sec

	Experimental	Standard	Porod
MW	70.433 kDa	75.814 kDa	-
Volume	-	-	105 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	323.9	1.8	321.37	2.21
Radius of gyration, Rg	3.02 nm	0.03	2.971 nm	0.031

	Min	Max
D	-	9.68
Guinier point	10	39