SASDF92: Urokinase plasminogen activator surface receptor, uPAR, T51C-V70C

基本情報

• 登録情報: データベース: SASBDB / ID: SASDF92

試料

Urokinase plasminogen activator surface receptor, uPAR, T51C-V70C

• Urokinase plasminogen activator surface receptor (protein), uPAR, Homo sapiens

機能・相同性

分子機能:

urokinase plasminogen activator receptor activity / Attachment of GPI anchor to uPAR / positive regulation of homotypic cell-cell adhesion / negative regulation of cysteine-type endopeptidase activity involved in apoptotic signaling pathway / urokinase plasminogen activator signaling pathway / regulation of plasminogen activation / regulation of fibrinolysis / protein complex involved in cell-matrix adhesion / regulation of proteolysis / serine-type endopeptidase complex / Dissolution of Fibrin Clot / positive regulation of epidermal growth factor receptor signaling pathway / extrinsic component of membrane / positive regulation of release of cytochrome c from mitochondria / positive regulation of DNA binding / regulation of cell adhesion / specific granule membrane / negative regulation of intrinsic apoptotic signaling pathway / cell projection / chemotaxis / blood coagulation / signaling receptor activity / positive regulation of protein phosphorylation / protein domain specific binding / endoplasmic reticulum lumen / external side of plasma membrane / focal adhesion / signaling receptor binding / Neutrophil degranulation / endoplasmic reticulum membrane / negative regulation of apoptotic process / enzyme binding / cell surface / signal transduction / extracellular region / membrane / plasma membrane

ドメイン・相同性:

CD59 antigen, conserved site / Ly-6 / u-PAR domain signature. / Ly-6 antigen / uPA receptor -like domain / u-PAR/Ly-6 domain / Ly-6 antigen/uPA receptor-like / Snake toxin-like superfamily

構成要素:

Urokinase plasminogen activator surface receptor

• 生物種: Homo sapiens (ヒト)
• 引用: ジャーナル: J Biol Chem / 年: 2019; タイトル: Did evolution create a flexible ligand-binding cavity in the urokinase receptor through deletion of a plesiotypic disulfide bond?; 著者: Julie M Leth / Haydyn D T Mertens / Katrine Zinck Leth-Espensen / Thomas J D Jørgensen / Michael Ploug / ; 要旨: The urokinase receptor (uPAR) is a founding member of a small protein family with multiple Ly6/uPAR (LU) domains. The motif defining these LU domains contains five plesiotypic disulfide bonds stabilizing its prototypical three-fingered fold having three protruding loops. Notwithstanding the detailed knowledge on structure-function relationships in uPAR, one puzzling enigma remains unexplored. Why does the first LU domain in uPAR (DI) lack one of its consensus disulfide bonds, when the absence of this particular disulfide bond impairs the correct folding of other single LU domain-containing proteins? Here, using a variety of contemporary biophysical methods, we found that reintroducing the two missing half-cystines in uPAR DI caused the spontaneous formation of the corresponding consensus 7-8 LU domain disulfide bond. Importantly, constraints due to this cross-link impaired (i) the binding of uPAR to its primary ligand urokinase and (ii) the flexible interdomain assembly of the three LU domains in uPAR. We conclude that the evolutionary deletion of this particular disulfide bond in uPAR DI may have enabled the assembly of a high-affinity urokinase-binding cavity involving all three LU domains in uPAR. Of note, an analogous neofunctionalization occurred in snake venom α-neurotoxins upon loss of another pair of the plesiotypic LU domain half-cystines. In summary, elimination of the 7-8 consensus disulfide bond in the first LU domain of uPAR have significant functional and structural consequences.

登録者

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

モデル

• モデル #2808: ; タイプ: dummy / ダミー原子の半径: 2.50 A; コメント: Refined DAMMIN model from average volume (10 DAMMIF iterations); カイ2乗値: 0.955 / P-value: 0.891460; 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: Urokinase plasminogen activator surface receptor, uPAR, T51C-V70C; 試料濃度: 0.30-2.80
• バッファ: 名称: 20 mM PBS, 5 %(v/v) glycerol / pH: 7.4

要素 #66

名称: uPAR / タイプ: protein / 記述: Urokinase plasminogen activator surface receptor / 分子量: 36.98 / 分子数: 1 / 由来: Homo sapiens / 参照: UniProt: Q03405
配列:

MGHPPLLPLL LLLHTCVPAS WGLRCMQCKT NGDCRVEECA LGQDLCRTTI VRLWEEGEEL ELVEKSCTHS EKTNRTLSYR TGLKITSLTE VVCGLDLCNQ GNSGRAVTYS RSRYLECISC GSSDMSCERG RHQSLQCRSP EEQCLDVVTH WIQEGEEGRP KDDRHLRGCG YLPGCPGSNG FHNNDTFHFL KCCNTTKCNE GPILELENLP QNGRQCYSCK GNSTHGCSSE ETFLIDCRGP MNQCLVATGT HEPKNQSYMV RGCATASMCQ HAHLGDAFSM NHIDVSCCTK SGCNHPDLDV QYRSGAAPQP GPAHLSLTIT LLMTARLWGG TLLWT

実験情報

• ビーム: 設備名称: PETRA III EMBL P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.124 Å / スペクトロメータ・検出器間距離: 3.1 mm
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: Urokinase plasminogen activator surface receptor, uPAR, T51C-V70C
測定日: 2017年12月1日 / 保管温度: 10 °C / セル温度: 10 °C / 照射時間: 0.05 sec. / フレーム数: 20 / 単位: 1/nm /

	Min	Max
Q	0.0725	5.0879

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 1125 /

	Min	Max
Q	0.072479	3.18188
P(R) point	1	1125
R	0	8.9

結果

カーブのタイプ: merged
コメント: Introduced disulfide in the first LU domain, DI, of uPAR (T51C-V70C).

	実験値	Standard	Standard error	Porod
分子量	46.108 kDa	46.108 kDa	5	32 kDa
体積	-	-	-	54.5 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	0.03608	0.004	0.036	0.001
慣性半径, Rg	2.592 nm	0.02	2.52 nm	0.01

	Min	Max	Error
D	-	8.9	0.5
Guinier point	1	161	-