UBE1 [6His-tagged], Preis auf Anfrage

Produktübersicht

• Artikelname: UBE1 [6His-tagged], Preis auf Anfrage
• Artikelnummer: UBI-61-0001-001
• Hersteller Artikelnummer: 61-0001-001
• Hersteller: Ubiquigent

Zusatzinformationen

• Species

  Human

• Source

  Sf21 insect cell-baculovirus expression

• Quantity

  10 μg

• Storage

  -70°C

• Concentration

  0.5 mg/ml

• Formulation

  50 mM HEPES pH 7.5 (NaOH), 150 mM NaCl, 2 mM dithiothreitol, 10% glycerol

• Molecular Weight

  ~121 kDa

• Stability

  12 months at -70°C; aliquot as required

• Protein Sequence

  Accession number: NP_003325. For full protein sequence information download the Certificate of Analysis pdf.

• QA; Protein Identification

  Confirmed by mass spectrometry.

• QA; Activity

  E1-Ubiquitin Thioester Loading Assay: The activity of His-UBE1 was validated by loading ubiquitin onto the active cysteine of His-UBE1. Incubation of the His-UBE1 enzyme in the presence of ubiquitin and ATP at 30oC was compared at two time points, T0 and T10 minutes. Sensitivity of the ubiquitin/His-UBE1 thioester bond to the reducing agent DTT was confirmed.

• Background

The enzymes of the ubiquitylation pathway play a pivotal role in a number of cellular processes including regulated and targeted proteasomal degradation of substrate proteins. Three classes of enzymes are involved in the process of ubiquitylation; activating enzymes (E1s), conjugating enzymes (E2s) and protein ligases (E3s). UBE1 is a member of the E1 activating enzyme family and cloning of the human gene was first described by Handley et al. (1991). The UBE1 gene has been mapped to Xp11.3-p11.23 by highresolution fluorescence in situ hybridization (Takahashi et al., 1992). UBE1 ‘activates’ ubiquitin through catalysing a C-terminal ATP dependent adenylation of the protein which results in it forming a high-energy thioester bond with the sulfhydryl group of UBE1. UBE1 is monomeric and there are two active sites within the UBE1 protein allowing it to bind two ubiquitin moieties at a time, with a new ubiquitin forming an adenylate intermediate as the previous one is transferred to the thiol site (Jin et al., 2007; Zheng et al., 2009). Defects in UBE1 are known to cause spinal muscular atrophy X-linked type 2 (SMAX2) also known as X-linked lethal infantile spinal muscular atrophy, distal X-linked arthrogryposis multiplex congenita or X-linked arthrogryposis type 1 (AMCX1). Spinal muscular atrophy refers to a group of neuromuscular disorders characterized by degeneration of the anterior horn cells of the spinal cord, leading to symmetrical muscle weakness and atrophy. SMAX2 is a lethal infantile form presenting with hypotonia, areflexia, and multiple congenital contractures (Ramser et al., 2008).

References:

Handley PM, Mueckler M, Siegel NR, Ciechanover A, Schwartz AL (1991) Molecular cloning, sequence, and tissue distribution of the human ubiquitin-activating enzyme E1. Proc Natl Acad Sci USA 88, 258-62.

Jin J, Li X, Gygi SP, Harper JW (2007) Dual E1 activation systems for ubiquitin differentially regulate E2 enzyme charging. Nature 447, 1135-8.

Ramser J, Ahearn ME, Lenski C, Yariz KO, Hellebrand H, von Rhein M, Clark RD, Schmutzler RK, Lichtner P, Hoffman EP, Meindl A, Baumbach-Reardon L. (2008) Rare missense and synonymous variants in UBE1 are associated with X-linked infantile spinal muscular atrophy. Am J Hum Genet 82, 188-93.

Takahashi E, Ayusawa D, Kaneda S, Itoh Y, Seno T, Hori T (1992) The human ubiquitin-activating enzyme E1 gene (UBE1) mapped to band Xp11.3—-p11.23 by fluorescence in situ hybridization. Cytogenet Cell Genet 59, 268-9.

Zheng M, Liu J, Yang Z, Gu X, Li F, Lou T, Ji C, Mao Y (2009) Expression, purification and characterization of human ubiquitin-activating enzyme, UBE1. Mol Biol Rep 37, 1413-9.