SASDCC9: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 42% D2O SANS

Basic information

Entry

Database: SASBDB / ID: SASDCC9

Sample

Nanolipoprotein Particle (NLP) from In Vitro Assembly - 42% D2O SANS

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

Function / homology

Function:

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 / Heme signaling / lipase inhibitor activity / Retinoid metabolism and transport / high-density lipoprotein particle receptor binding / spherical high-density lipoprotein particle / regulation of intestinal cholesterol absorption / protein oxidation / 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 / HDL assembly / high-density lipoprotein particle binding / 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 / Platelet degranulation / discoidal high-density lipoprotein particle / peptidyl-methionine modification / intermediate-density lipoprotein particle / phosphatidylcholine biosynthetic process / glucocorticoid metabolic process / acylglycerol homeostasis / phosphatidylcholine-sterol O-acyltransferase activator activity / positive regulation of phospholipid efflux / positive regulation of cholesterol metabolic process / lipid storage / phospholipid homeostasis / lipid transporter activity / chylomicron / high-density lipoprotein particle remodeling / phospholipid efflux / phospholipid transport / chemorepellent activity / cholesterol transfer activity / reverse cholesterol transport / very-low-density lipoprotein particle / high-density lipoprotein particle assembly / low-density lipoprotein particle / lipoprotein biosynthetic process / cholesterol transport / high-density lipoprotein particle / regulation of Cdc42 protein signal transduction / triglyceride homeostasis / endothelial cell proliferation / negative regulation of interleukin-1 beta production / cholesterol efflux / positive regulation of Rho protein signal transduction / adrenal gland development / cholesterol binding / 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 / positive regulation of stress fiber assembly / heat shock protein binding / 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

Domain/homology:

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

Component:

Apolipoprotein A-I

• Biological species: Mus musculus (house mouse)
• Citation: Journal: Protein Sci / Year: 2018; Title: Small-angle X-ray and neutron scattering demonstrates that cell-free expression produces properly formed disc-shaped nanolipoprotein particles.; Authors: Thomas E Cleveland / Wei He / Angela C Evans / Nicholas O Fischer / Edmond Y Lau / Matthew A Coleman / Paul Butler / ; Abstract: 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.

Contact author

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

Models

Sample

• Sample: Name: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 42% D2O SANS; Specimen concentration: 0.96 mg/ml / Entity id: 861 / 862
• Buffer: Name: PBS in 42% D2O / pH: 7.4 ; Comment: 8 g/L NaCl, 0.2 g/L KH2PO4, 1.15 g/L Na2HPO4, 0.2 g/L KCl, in 42% D2O with no further pH adjustment

Entity #861

Name: ApoA-I / Type: protein / Description: Apolipoprotein A-I / Formula weight: 25.153 / Num. of mol.: 2 / Source: Mus musculus / References: UniProt: Q00623
Sequence:

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

• Entity #862: Name: DMPC / Type: other / Description: 1,2-dimyristoyl-sn-glycero-3-phosphocholine / Formula weight: 0.678

Experimental information

• Beam: Instrument name: NIST Center for Neutron Research NGB30 / City: Gaithersburg, MD / 国: USA / Type of source: neutron source / Wavelength: 0.6 Å / Dist. spec. to detc.: 4 mm
• Detector: Name: Ordela 2660N / Type: 640x640 mm 3He posn sensitive prop counter / Pixsize x: 5.08 mm

Scan

Title: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 42% D2O SANS
Measurement date: Dec 11, 2015 / Storage temperature: 4 °C / Cell temperature: 25 °C / Exposure time: 3600 sec. / Number of frames: 2 / Unit: 1/A /

	Min	Max
Q	0.01	0.4768

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 95 /

	Min	Max
Q	0.009974	0.2747
P(R) point	1	95
R	0	72.73

Result

Experimental MW: 142 kDa / Type of curve: single_conc
Comments: Additional SANS measurement parameters. Two detector positions (in m) and count times (s) were employed: 1.33 m (two frames of 900 + 831 = 1731 s total count time); 4 m (two frames of 3600 + 3600 = 7200 s total count time).

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	0.043	0.001	0.044	0.00084
Radius of gyration, Rg	2.87 nm	0.06	2.91 nm	0.19

	Min	Max
D	-	7.27
Guinier point	1	28