Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker.

Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker. Research Abstract Details 

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Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker. Abstract Text:

Brett Feeney,Erik J Soderblom,Michael B Goshe,A Clay Clark,

Coupled with over-expression in host organisms, fusion protein systems afford economical methods to obtain large quantities of target proteins in a fast and efficient manner. Some proteases used for these purposes cleave C-terminal to their recognition sequences and do not leave extra amino acids on the target. However, they are often inefficient and are frequently promiscuous, resulting in non-specific cleavages of the target protein. To address these issues, we created a fusion protein system that utilizes a highly efficient enzyme and leaves no residual amino acids on the target protein after removal of the affinity tag. We designed a glutathione S-transferase (GST)-fusion protein vector with a caspase-3 consensus cleavage sequence located between the N-terminal GST tag and a target protein. We show that the enzyme efficiently cleaves the fusion protein without leaving excess amino acids on the target protein. In addition, we used an engineered caspase-3 enzyme that is highly stable, has increased activity relative to the wild-type enzyme, and contains a poly-histidine tag that allows for efficient removal of the enzyme after cleavage of the fusion protein. Although we have developed this system using a GST tag, the system is amenable to any commercially available affinity tag.

Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker. Publishing Authors By Initials

B Feeney,EJ Soderblom,MB Goshe,AC Clark,

For similar proteins: recombinant proteins: recombinant fusion proteins research abstracts see: proteins: recombinant proteins: recombinant fusion proteins research

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Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Protein expression and purification

VOLUME: 47

Page Numbers: 311-8

Journal Abbreviation: Protein Expr. Purif.

ISSN: 1046-5928

DAY: 27

MONTH: 10

YEAR: 2005

Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 9101496

Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker. Keywords Mesh Terms:

KEYWORDS: Recombinant Fusion Proteins

MESH TERMS: metabolism

Chemical & Substance for Abstract: Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker. Information

Substance Name: Caspase 3

Registry Number: EC 3.4.22.-

Grant and Affiliation Information for Novel protein purification system utilizing an N-terminal fusion protein and a caspase-3 cleavable linker.

AFFILIATION: Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695, USA.

Country: United States

AGENCY: United States NIGMS

GRANT: GM065970

ACRONYM: GM

MEDLINETA: Protein Expr Purif

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