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

基本情報

• 登録情報: データベース: SASBDB / ID: SASDDS4

試料

cGMP-dependent protein kinase 1: ∆53 PKG Iα

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

機能・相同性

分子機能:

cGMP effects / cGMP-dependent protein kinase / cGMP-dependent protein kinase activity / Rap1 signalling / negative regulation of platelet aggregation / regulation of GTPase activity / cGMP binding / calcium channel regulator activity / protein serine kinase activity / signal transduction / ATP binding / identical protein binding / cytosol / cytoplasm

ドメイン・相同性:

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

構成要素:

cGMP-dependent protein kinase 1

• 生物種: Bos taurus (ウシ)
• 引用: ジャーナル: J Biol Chem / 年: 2018; タイトル: An N-terminally truncated form of cyclic GMP-dependent protein kinase Iα (PKG Iα) is monomeric and autoinhibited and provides a model for activation.; 著者: Thomas M Moon / Jessica L Sheehe / Praveena Nukareddy / Lydia W Nausch / Jessica Wohlfahrt / Dwight E Matthews / Donald K Blumenthal / Wolfgang R Dostmann / ; 要旨: 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.

登録者

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

モデル

• モデル #1814: ; タイプ: dummy / ソフトウェア: (5.0) / ダミー原子の半径: 3.25 A / 対称性: p1 / コメント: Filtered, Averaged 3-D envelope / カイ2乗値: 0.602 / P-value: 0.999000; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: cGMP-dependent protein kinase 1: ∆53 PKG Iα
• バッファ: 名称: 50 mM MES, 300 mM NaCl, 1 mM TCEP, 5 mM DTT / pH: 6.9

要素 #988

名称: ∆53 PKG Iα / タイプ: protein / 記述: cGMP-dependent protein kinase 1 / 分子量: 70.433 / 分子数: 1 / 由来: Bos taurus / 参照: UniProt: P00516
配列:

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

実験情報

• ビーム: 設備名称: Stanford Synchrotron Radiation Lightsource (SSRL) BL4-2; 地域: Stanford, CA / 国: USA / 線源: X-ray synchrotron / 波長: 0.1127 Å / スペクトロメータ・検出器間距離: 1.7 mm
• 検出器: 名称: Rayonix MX225-HE

スキャン

タイトル: cGMP-dependent protein kinase 1: ∆53 PKG Iα
測定日: 2015年6月6日 / 保管温度: 4 °C / セル温度: 22 °C / 照射時間: 1 sec. / フレーム数: 600 / 単位: 1/A /

	Min	Max
Q	0.0087	0.5136

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 192 /

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

結果

カーブのタイプ: sec

	実験値	Standard	Porod
分子量	70.433 kDa	75.814 kDa	-
体積	-	-	105 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	323.9	1.8	321.37	2.21
慣性半径, Rg	3.02 nm	0.03	2.971 nm	0.031

	Min	Max
D	-	9.68
Guinier point	10	39