Alpha-Synuclein, Recombinant, Human

Cosmo Bio

Catalog No.: CSR-SYN04-2

Product Description

E. coli expressed human α-synuclein has been widely used in structural and functional studies. Masuda et. al. (1)  have reported that approximately 20% of E. coli expressed human α-syn is mistranslated resulting in a Cys residue instead of Tyr at position 136 and causing an increase in the levels of dimeric α-synuclein.

CSR-SYN04 is E. coli human alpha-Synuclein (human) expressed from a plasmid in which the TAC codon coding for Tyr136 in the human protein is replaced by TAT to produce protein expressing Tyr at position 136 homogenously. It can be used as human α-syn under similar conditions to that of in vivo.

This product can also be used for comparison (control) of α-Synuclein Fibrils (CSR-SYN03).

Source: Human
Expression System: E.coli
Concentration: 1mg/mL
Purity: ≥ 90% (SDS-Page)
Storage: -20°C

References

(1) Masuda M, Dohmae N, Nonaka T, Oikawa T, Hisanaga S, Goedert M, Hasegawa M. Cysteine misincorporation in bacterially expressed human alpha-synuclein. FEBS Lett. 2006 Mar 20;580(7):1775-9. doi: 10.1016/j.febslet.2006.02.032. Epub 2006 Feb 24. PMID: 16513114.

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Alpha-Synuclein, Human	CSR-SYN04
Alpha-Synuclein Fibrils, Mouse	CSR-SYN05
Alpha-Synuclein, Recombinant, Mouse	CSR-SYN06

Alpha-Synuclein, Recombinant, Human FAQs

Has Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) been purified or filtered (e.g., by ultrafiltration)?
No. Recombinant Human Alpha-Synuclein (E. coli expressed) has not undergone filtration processes such as ultrafiltration.

Does Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) contain any tag sequences?
No. Recombinant Human Alpha-Synuclein (E. coli expressed) does not contain any tags.

What is the molecular weight of Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04)?
The molecular weight of Recombinant Human Alpha-Synuclein (E. coli expressed) is 14.5 kDa.

What are the amino acid sequences of Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) and Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04)?
Both products have the same amino acid sequence (140 amino acids):
MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAV
VTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA

Can I detect Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) and Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) using a phospho-specific antibody?
No. Recombinant Human Alpha-Synuclein (E. coli expressed) is not phosphorylated, and Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) are prepared by fibrillization of this non-phosphorylated protein. Therefore, neither product is expected to be detected by phospho-specific α-synuclein antibodies. If a signal is observed in Western blot analysis, it is likely due to non-specific cross-reactivity of the antibody.

How is Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) prepared?
The full-length Alpha-synuclein is expressed in E. coli. Monomers purified from the lysate are subjected to ultracentrifugation (160,000 × g for 20 min), and the resulting supernatant is designated as Recombinant Human Alpha-Synuclein (E. coli expressed).

Regarding codon optimization in Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04), what changes were made to the construct sequence?
The full-length Alpha-synuclein gene sequence was modified only at residue Tyr136, where the codon was changed from TAC to TAT. No other changes were introduced.
Sequence:
ATGGATGTATTCATGAAAGGACTTTCAAAGGCCAAGGAGGGAGTTGTGGCTGCTGCTG
AGAAAACCAAACAGGGTGTGGCAGAAGCAGCAGGAAAGACAAAAGAGGGTGTTCTCT
ATGTAGGCTCCAAAACCAAGGAGGGAGTGGTGCATGGTGTGGCAACAGTGGCTGAGA
AGACCAAAGAGCAAGTGACAAATGTTGGAGGAGCAGTGGTGACGGGTGTGACAGCAG
TAGCCCAGAAGACAGTGGAGGGAGCAGGGAGCATTGCAGCAGCCACTGGCTTTGTCA
AAAAGGACCAGTTGGGCAAGAATGAAGAAGGAGCCCCACAGGAAGGAATTCTGGAAG
ATATGCCTGTGGATCCTGACAATGAGGCTTATGAAATGCCTTCTGAGGAAGGGTATCAA
GACTATGAACCTGAAGCCTAA

Reference:
Cysteine misincorporation in bacterially expressed human α-synuclein
Masami Masuda et al. FEBS Lett. 2006 Mar 20;580(7):1775-9. doi: 10.1016/j.febslet.2006.02.032. Epub 2006 Feb 24.