SASDC99: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 100% D2O SANS

基本情報

登録情報

データベース: SASBDB / ID: SASDC99

試料

Nanolipoprotein Particle (NLP) from In Vitro Assembly - 100% D2O SANS

• Apolipoprotein A-I (protein), ApoA-I, Mus musculus
• 1,2-dimyristoyl-sn-glycero-3-phosphocholine (other), DMPC

機能・相同性

分子機能:

ABC transporters in lipid homeostasis / Chylomicron assembly / Chylomicron remodeling / HDL remodeling / Scavenging by Class B Receptors / HDL clearance / Scavenging of heme from plasma / Scavenging by Class A Receptors / lipase inhibitor activity / Retinoid metabolism and transport / Heme signaling / high-density lipoprotein particle receptor binding / spherical high-density lipoprotein particle / protein oxidation / regulation of intestinal cholesterol absorption / vitamin transport / Regulation of Insulin-like Growth Factor (IGF) transport and uptake by Insulin-like Growth Factor Binding Proteins (IGFBPs) / Post-translational protein phosphorylation / cholesterol import / high-density lipoprotein particle binding / HDL assembly / blood vessel endothelial cell migration / negative regulation of heterotypic cell-cell adhesion / apolipoprotein A-I receptor binding / negative regulation of cytokine production involved in immune response / negative regulation of cell adhesion molecule production / negative regulation of very-low-density lipoprotein particle remodeling / peptidyl-methionine modification / discoidal high-density lipoprotein particle / intermediate-density lipoprotein particle / phosphatidylcholine biosynthetic process / Platelet degranulation / glucocorticoid metabolic process / acylglycerol homeostasis / phosphatidylcholine-sterol O-acyltransferase activator activity / positive regulation of phospholipid efflux / positive regulation of cholesterol metabolic process / lipid storage / chylomicron / lipid transporter activity / phospholipid homeostasis / high-density lipoprotein particle remodeling / phospholipid efflux / phospholipid transport / cholesterol transfer activity / chemorepellent activity / reverse cholesterol transport / high-density lipoprotein particle assembly / cholesterol transport / very-low-density lipoprotein particle / low-density lipoprotein particle / lipoprotein biosynthetic process / high-density lipoprotein particle / regulation of Cdc42 protein signal transduction / triglyceride homeostasis / endothelial cell proliferation / cholesterol efflux / negative regulation of interleukin-1 beta production / cholesterol binding / adrenal gland development / positive regulation of Rho protein signal transduction / cholesterol biosynthetic process / lipoprotein particle binding / endocytic vesicle / negative regulation of tumor necrosis factor-mediated signaling pathway / positive regulation of cholesterol efflux / animal organ regeneration / phospholipid metabolic process / positive regulation of substrate adhesion-dependent cell spreading / positive regulation of phagocytosis / heat shock protein binding / positive regulation of stress fiber assembly / cholesterol metabolic process / cholesterol homeostasis / integrin-mediated signaling pathway / regulation of protein phosphorylation / phospholipid binding / negative regulation of inflammatory response / extracellular vesicle / amyloid-beta binding / protein stabilization / G protein-coupled receptor signaling pathway / lipid binding / enzyme binding / cell surface / protein homodimerization activity / extracellular space / extracellular region / identical protein binding / cytosol

ドメイン・相同性:

Apolipoprotein A/E / : / Apolipoprotein A1/A4/E domain

構成要素:

Apolipoprotein A-I

• 生物種: Mus musculus (ハツカネズミ)
• 引用: ジャーナル: Protein Sci / 年: 2018; タイトル: Small-angle X-ray and neutron scattering demonstrates that cell-free expression produces properly formed disc-shaped nanolipoprotein particles.; 著者: Thomas E Cleveland / Wei He / Angela C Evans / Nicholas O Fischer / Edmond Y Lau / Matthew A Coleman / Paul Butler / ; 要旨: Nanolipoprotein particles (NLPs), composed of membrane scaffold proteins and lipids, have been used to support membrane proteins in a native-like bilayer environment for biochemical and structural studies. Traditionally, these NLPs have been prepared by the controlled removal of detergent from a detergent-solubilized protein-lipid mixture. Recently, an alternative method has been developed using direct cell-free expression of the membrane scaffold protein in the presence of preformed lipid vesicles, which spontaneously produces NLPs without the need for detergent at any stage. Using SANS/SAXS, we show here that NLPs produced by this cell-free expression method are structurally indistinguishable from those produced using detergent removal methodologies. This further supports the utility of single step cell-free methods for the production of lipid binding proteins. In addition, detailed structural information describing these NLPs can be obtained by fitting a capped core-shell cylinder type model to all SANS/SAXS data simultaneously.

登録者

• Thomas Cleveland (National Institute of Standards and Technology, Gaithersburg, MD, USA)

モデル

試料

• 試料: 名称: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 100% D2O SANS; 試料濃度: 5.21 mg/ml / Entity id: 861 / 862
• バッファ: 名称: PBS in D2O / pH: 7.4 ; コメント: 8 g/L NaCl, 0.2 g/L KH2PO4, 1.15 g/L Na2HPO4, 0.2 g/L KCl, in D2O with no additional pH adjustment

要素 #861

名称: ApoA-I / タイプ: protein / 記述: Apolipoprotein A-I / 分子量: 25.153 / 分子数: 2 / 由来: Mus musculus / 参照: UniProt: Q00623
配列:

MHHHHHHGEN LYFQGMLNLL ENWDTLGSTV SQLQERLGPL TRDFWDNLEK ETDWVRQEMN KDLEEVKQKV QPYLDEFQKK WKEDVELYRQ KVAPLGAELQ ESARQKLQEL QGRLSPVAEE FRDRMRTHVD SLRTQLAPHS EQMRESLAQR LAELKSNPTL NEYHTRAKTH LKTLGEKARP ALEDLRHSLM PMLETLKTKA QSVIDKASET LTAQ

• 要素 #862: 名称: DMPC / タイプ: other / 記述: 1,2-dimyristoyl-sn-glycero-3-phosphocholine / 分子量: 0.678

実験情報

• ビーム: 設備名称: NIST Center for Neutron Research NG7 / 地域: Gaithersburg, MD / 国: USA / 線源: neutron source / 波長: 0.6 Å / スペクトロメータ・検出器間距離: 4 mm
• 検出器: 名称: Ordela 2660N / タイプ: 640x640 mm 3He posn sensitive prop counter / Pixsize x: 5.08 mm

スキャン

タイトル: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 100% D2O SANS
測定日: 2015年11月25日 / 保管温度: 4 °C / セル温度: 20 °C / 照射時間: 1800 sec. / フレーム数: 2 / 単位: 1/A /

	Min	Max
Q	0.0039	0.5936

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 161 /

	Min	Max
Q	0.004706	0.2573
P(R) point	1	161
R	0	90.92

結果

Experimental MW: 142 kDa / カーブのタイプ: single_conc
コメント: Additional SANS measurement parameters. Three detector positions (in m) and count times (s) were employed: 1 m (two frames of 300 + 900 = 1200 s total count time); 4 m (two frames of 1800 + 1800 = 3600 s total count time); 13 m (two frames of 300 + 600 = 900 s total count time).

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	2.621	0.006	2.6	0.0046
慣性半径, Rg	3.121 nm	0.007	3.11 nm	0.06

	Min	Max
D	-	9.09
Guinier point	3	86