Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
PRO-404

Background:

Erythrocyte spectrin is the main component of the red cell membrane skeleton responsible for the shape and physical properties of red cells. Spectrin is composed of 2 subunits alpha and beta (280 and 246 kDa respectively). The human erithroid a-spectrin (2418 amino acids) consists of 22 repeating segments of about 106 amino acids in length. This protein was demonstrated to be
a specific substrate for ubiquitina tion in vitro and in vivo. The main ubiquitination site on human
a-spectrin was identified in the lysine 27 of the repeating segment 17 by sitedirected mutagenesis. Cell-free experiments using radiolabeled, biotinylated or native ubiquitin and cellular lysates from rabbit retyculocytes, human erythrocytes or k562 cells, showed that only monoubiquitination occurs at this site.

Description :

Recombinant fusion protein: Glutathione-S-Transferase (GST) and a fragment of human a-spectrin (from Lysine 1601 to Leucine 1818) containing repeat 17 and the ubiquitination target lysine 27.
The total molecular weight of the fusion protein is 52.4 kDa.

Presentation:

Sterile Filtered clear solution.

Formulation:

The protein (1.7mg/ml) contains 50mM Tris buffer pH-8 and 10mM glutathione.

Stability:

rGST-alpha Spectrin although stable at 14⁰C for 1 week, should be stored desiccated below -18⁰C.
Please prevent freeze-thaw cycles.

Purity:

Greater than 90.0% as determined by both:
(a) Analysis by RP-HPLC.
(b) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel

Application:

Substrate for cell-free ubiquitination assays.

References:

1. Speicher D.W., Morrow J.S., Knowles W.J., Marchesi V.T. (1982) J. Biol. Chem. 257, 9093-9101
2. Corsi D., Galluzzi L., Crinelli R., Magnani M. (1995) J. Biol. Chem. 270, 8928-8935
3. Galluzzi L., Paiardini M., Lecomte M.C., Magnani M. (2001) FEBS Letters 489, 254-258

Legal Consideration:

This material is offered by ProSpec-TechnoGene for research, laboratory or further manufacturing purposes.

Gene:

Name:SPTA1
Synonyms:SPTA

Protein synonyms/aliases:

Spectrin alpha chain, erythrocyte (Erythroid alpha-spectrin).

Protein Family:

Belongs to the spectrin family.

Protein Domains:

Domains:
•IPR001452   Src homology-3
•IPR002017   Spectrin repeat
•IPR002048   Calcium-binding EF-hand

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. A Single Mutation Induces Amyloid Aggregation in the alpha-Spectrin SH3 Domain: Analysis of the Early Stages of Fibril Formation.
J Mol Biol 2 006 Feb 17;356(2):453-68
2. A novel splicing mutation of the alpha-spectrin gene in the original hereditary pyropoikilocytosis kindred.
Blood 2005 Dec 15;106(13):4367-9
3. Detection of dynamic water molecules in a microcrystalline sample of the SH3 domain of alpha-spectrin by MAS solid-state N MR.
J Biomol NMR 2005 Apr;31(4):295-310
4. Mutational effects at the tetramerization site of nonerythroid alpha spectrin.
Brain Res Mol Brain Res 2005 May 20;136(1-2):81-90
5. Ubiquitination of red blood cell alpha-spectrin does not affect heterodimer formation.
Am J Hematol 2005 Apr;78(4):281-7
6. Pseudohypoaldosteronism type 1 and the genes encoding prostasin, alpha-spectrin, and Nedd4.
Int J Mol Med 2004 Dec;14(6):1101-4

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中華民國95年06月06日更新