SASDAH4: DH-PH (DH-PH module of PDZRhoGEF)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDAH4

試料

DH-PH

• DH-PH module of PDZRhoGEF (protein), DH-PH, Homo sapiens

• 生物種: Homo sapiens (ヒト)
• 引用: ジャーナル: Protein Sci / 年: 2009; タイトル: The solution structure and dynamics of the DH-PH module of PDZRhoGEF in isolation and in complex with nucleotide-free RhoA.; 著者: Tomasz Cierpicki / Jakub Bielnicki / Meiying Zheng / Jakub Gruszczyk / Marta Kasterka / Maxim Petoukhov / Aming Zhang / Erik J Fernandez / Dmitri I Svergun / Urszula Derewenda / John H Bushweller / Zygmunt S Derewenda / ; 要旨: The DH-PH domain tandems of Dbl-homology guanine nucleotide exchange factors catalyze the exchange of GTP for GDP in Rho-family GTPases, and thus initiate a wide variety of cellular signaling cascades. Although several crystal structures of complexes of DH-PH tandems with cognate, nucleotide free Rho GTPases are known, they provide limited information about the dynamics of the complex and it is not clear how accurately they represent the structures in solution. We used a complementary combination of nuclear magnetic resonance (NMR), small-angle X-ray scattering (SAXS), and hydrogen-deuterium exchange mass spectrometry (DXMS) to study the solution structure and dynamics of the DH-PH tandem of RhoA-specific exchange factor PDZRhoGEF, both in isolation and in complex with nucleotide free RhoA. We show that in solution the DH-PH tandem behaves as a rigid entity and that the mutual disposition of the DH and PH domains remains identical within experimental error to that seen in the crystal structure of the complex, thus validating the latter as an accurate model of the complex in vivo. We also show that the nucleotide-free RhoA exhibits elevated dynamics when in complex with DH-PH, a phenomenon not observed in the crystal structure, presumably due to the restraining effects of crystal contacts. The complex is readily and rapidly dissociated in the presence of both GDP and GTP nucleotides, with no evidence of intermediate ternary complexes.

登録者

• Maxim Petoukhov (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

モデル

• モデル #61: ; タイプ: dummy / ソフトウェア: Dammin / ダミー原子の半径: 1.90 A / カイ2乗値: 2.458624; Omokage検索でこの集合体の類似形状データを探す (詳細)
• モデル #62: ; タイプ: atomic / ソフトウェア: Crysol / カイ2乗値: 2.399401; 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: DH-PH / Sample MW: 41 kDa / 試料濃度: 0.86-4.70
• バッファ: 名称: 50 mM Tris-HCL / pH: 7.5 / 組成: 150 mM NaCl, 1.0 mM DTT

要素 #57

名称: DH-PH / タイプ: protein / 記述: DH-PH module of PDZRhoGEF / 分子量: 41 / 分子数: 1 / 由来: Homo sapiens
配列:

QNWQHTVGKD VVAGLTQREI DRQEVINELF VTEASHLRTL RVLDLIFYQR MKKENLMPRE ELARLFPNLP ELIEIHNSWC EAMKKLREEG PIIKEISDLM LARFDGPARE ELQQVAAQFC SYQSIALELI KTKQRKESRF QLFMQEAESH PQCRRLQLRD LIISEMQRLT KYPLLLESII KHTEGGTSEH EKLCRARDQC REILKYVNEA VKQTENRHRL EGYQKRLDAT ALERASNPLA AEFKSLDLTT RKMIHEGPLT WRISKDKTLD LHVLLLEDLL VLLQKQDEKL LLKCTFSPVL KLNAVLIRSV ATDKRAFFII CTSKLGPPQI YELVALTSSD KNTWMELLEE AVRNA

実験情報

• ビーム: 設備名称: DORIS III X33 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron
• 検出器: 名称: MAR 345 Image Plate

スキャン

タイトル: DH-PH / 測定日: 2008年10月30日 / 保管温度: 10 °C / セル温度: 10 °C / 単位: 1/nm /

	Min	Max
Q	0.2295	4.816

距離分布関数 P(R)

ソフトウェア P(R): GNOM 4.5a / ポイント数: 369 /

	Min	Max
Q	0.2571	2.796
P(R) point	12	380
R	0	8.941

結果

I0 from Guinier: 0.378 / Rg from Guinier: 2.9 nm / カーブのタイプ: merged / Standard: BSA /

	実験値	Porod
分子量	40 kDa	-
体積	-	60 nm3

/

	Min	Max
D	-	9
Guinier point	1	96