M7074

Membrane Scaffold Protein 1E3D1

recombinant, expressed in E. coli

• Synonym(s): MSP1E3D1
• NACRES: NA.26
• UNSPSC Code: 12352202
• Form: lyophilized powder
• Biological source: Streptomyces kanamyceticus
• Recombinant: expressed in E. coli
• Mol wt: M_w 32599.98 by amino acid sequence

Properties

• biological source: Streptomyces kanamyceticus
• Quality Level: 200
• recombinant: expressed in E. coli
• description: N-Terminal histidine-tagged
• form: lyophilized powder
• mol wt: M_w 32599.98 by amino acid sequence
• ε (extinction coefficient): 26,600 M^-1cm^-1 at 280 nm (His-tag-cleaved dissolved in 20 mM Tris pH 7.4, 0.1M NaCl, 0.5mM EDTA and 0.01%NaN₃)(lit.), 29,400 M^-1cm^-1 at 280 nm (uncleaved His-tagged dissolved in 20 mM Tris pH 7.4, 0.1M NaCl, 0.5mM EDTA and 0.01%NaN₃)(lit.)
• storage temp.: −20°C

Description

General description

Sequence: GHHHHHHHDYDIPTTENLYFQGSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKVQPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEMRDRARAHVDALRTHLAPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEMRDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEYHAKATEHLSTLSEKAKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQ

The nanodisc concept is derived from high density lipoprotein (HDL) particles and their primary protein component, apolipoprotein. The nanodisc is a non-covalent structure of phospholipid bilayer and membrane scaffold protein (MSP), a genetically engineered protein, which mimics the function of Apolipoprotein A-1 (ApoA-1). A soluble nanodisc assembles as the phospholipid forms a bilayer, which is encircled by two amphipathic MSP molecules covering the hydrophobic alkyl chains of the bilayer. The length of the MSP controls the size of the nanodisc structure. MSP1E3D1 yields nanodiscs of ~12.9 nm. The thickness of a nanodisc is dependent on the type of phospholipid incorporated (typically 4.6−5.6 nm).

Application

For an extensive list of citations and protocols visit the Sligar Lab Website at; sligarlab.life.uiuc.edu/nanodisc.html

For guidelines on the use of this and other MSP′s to prepare Nanodiscs, please visit our Protocols for Membrane Scaffold Proteins and Nanodisc Formation page.

Nanodisc soluble lipid bilayer systems have proven to be a widely applicable means for rendering membrane proteins soluble in aqueous solutions in a native-like bilayer environment where they remain monodisperse and active. The critical component of nanodiscs is the encircling amphipathic helical protein belt (membrane scaffold protein).

The nanodisc system has been employed to incorporate a wide variety of proteins including GPCRs, P450s, bacteriorhodopsin, coagulation factors, cholera toxin, TAR receptor and aromatase.

Biochem/physiol Actions

Generates Nanodiscs ~12.9 nm in diameter

Physical form

Supplied as a lyophilized histidine-tagged protein with a TEV protease cleavage site stabilized with Tris-HCl, EDTA, and NaCl.

Legal Information

Nanodisc technology, and many of its uses, are covered by the following patents held by the University of Illinois.

• 7,691,414 Membrane scaffold proteins
• 7,662,410 Membrane scaffold proteins and embedded membrane proteins
• 7,622,437 Tissue factor compositions and methods
• 7,592,008 Membrane scaffold proteins
• 7,575,763 Membrane scaffold proteins and tethered membrane proteins
• 7,083,958 Membrane scaffold proteins
• 7,048,949 Membrane scaffold proteins

Safety Information

• pictograms: GHS07
• signalword: Warning
• hcodes: H315,H319,H335
• pcodes: P261 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Skin Irrit. 2 - STOT SE 3
• target_organs: Respiratory system
• Storage Class: 11 - Combustible Solids
• wgk: WGK 3
• flash_point_f: Not applicable
• flash_point_c: Not applicable