Avoiding Proteolysis During Protein Purification.
B. J. Ryan, G. Henehan
Published 2017 · Chemistry, Medicine
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All cells contain proteases which hydrolyze the peptide bonds between amino acids in a protein backbone. Typically, proteases are prevented from nonspecific proteolysis by regulation and by their physical separation into different subcellular compartments; however, this segregation is not retained during cell lysis, which is the initial step in any protein isolation procedure. Prevention of proteolysis during protein purification often takes the form of a two-pronged approach; firstly inhibition of proteolysis in situ, followed by the early separation of the protease from the protein of interest via chromatographical purification. Protease inhibitors are routinely used to limit the effect of the proteases before they are physically separated from the protein of interest via column chromatography. Here, commonly used approaches to reducing or avoiding proteolysis during protein purification and subsequent chromatography are reviewed.
This paper references
Sequence, expression and modeled structure of an aspartic proteinase from the human malaria parasite Plasmodium falciparum.
J. B. Dame (1994)
New approaches for dissecting protease functions to improve probe development and drug discovery
Edgar Deu (2012)
 Families of serine peptidases
N. Rawlings (1994)
Methods for samples preparation in proteomic research.
A. Bodzoń-Kułakowska (2007)
Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems
K. Terpe (2006)
Review: novel cysteine proteases of the papain family.
F. Bühling (2000)
Protease proteomics: revealing protease in vivo functions using systems biology approaches.
A. Doucet (2008)
In-gel nonspecific proteolysis for elucidating glycoproteins: a method for targeted protein-specific glycosylation analysis in complex protein mixtures.
Charles C Nwosu (2013)
An efficient method for purification of nonspecific lipid transfer protein-1 from rice seeds using kiwifruit actinidin proteolysis and ion exchange chromatography.
Sirous Ghobadi (2012)
Characterization of a PRL protein tyrosine phosphatase from Plasmodium falciparum.
P. R. Pendyala (2008)
Overview of Approaches to Preventing and Avoiding Proteolysis During Expression and Purification of Proteins
B. J. Ryan (2013)
Blunting Nature's Swiss Army Knife
C. Seife (1997)
The ADAM metalloproteinases
D. Edwards (2008)
Measurement of alkaline phosphatase of intestinal origin in plasma by p-bromotetramisole inhibition.
T. Kuwana (1991)
Interactions between human glutamate carboxypeptidase II and urea-based inhibitors: structural characterization.
C. Bařinka (2008)
Inhibition of unwanted proteolysis during sample preparation: Evaluation of its efficiency in challenge experiments
L. Castellanos-Serra (2002)
Identification and quantification of Fc fusion peptibody degradations by limited proteolysis method.
Lei Yu (2012)
Zymography methods for visualizing hydrolytic enzymes
Jennifer Vandooren (2013)
Stabilisation of Recombinant Aequorin by Polyols: Activity, Thermostability and Limited Proteolysis
M. Zeinoddini (2013)
Glyco-analytical multispecific proteolysis (Glyco-AMP): a simple method for detailed and quantitative Glycoproteomic characterization.
Serenus Hua (2013)
A biotechnology perspective of fungal proteases
Paula Monteiro de Souza (2015)
Activity‐Based Probes for the Study of Proteases: Recent Advances and Developments
Sevnur Serim (2012)
Mammalian cell protein expression for biopharmaceutical production.
J. Zhu (2012)
The Threonine Protease Activity of Testes-Specific Protease 50 (TSP50) Is Essential for Its Function in Cell Proliferation
Yuyin Li (2012)
Recombinant protein production in yeasts.
D. Mattanovich (2012)
The Hallmarks of Cancer
D. Hanahan (2000)
Asparagine Peptide Lyases
N. Rawlings (2011)
Bioinformatic Approaches for Predicting substrates of Proteases
Jiangning Song (2011)
Handbook of Enzyme Inhibitors
H. Zollner (1989)
MEROPS: the peptidase database
N. Rawlings (2010)
Molecular mechanisms of the protein serine/threonine phosphatases.
D. Barford (1996)
Prevention of unwanted proteolysis.
R. Beynon (1988)
Demonstration that 1-trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64) is one of the most effective low Mr inhibitors of trypsin-catalysed hydrolysis. Characterization by kinetic analysis and by energy minimization and molecular dynamics simulation of the E-64-beta-trypsin complex.
S. Sreedharan (1996)
The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La.
C. Chung (1981)
Bacterial expression systems for recombinant protein production: E. coli and beyond.
R. Chen (2012)
A simple fluorescence labeling method for studies of protein oxidation, protein modification, and proteolysis.
Andrew M Pickering (2012)
Lon and ClpP proteases participate in the physiological disintegration of bacterial inclusion bodies.
A. Vera (2005)
Detection of peptidases in Trypanosoma cruzi epimastigotes using chromogenic and fluorogenic substrates.
N. Healy (1992)
Proteases in Cellular Regulation Minireview Series*
T. Vanaman (1999)
Use of vanadate as protein-phosphotyrosine phosphatase inhibitor.
J. Gordon (1991)
ATP-dependent degradation of ubiquitin-protein conjugates.
A. Hershko (1984)
Reaction of serine proteases with substituted isocoumarins: discovery of 3,4-dichloroisocoumarin, a new general mechanism based serine protease inhibitor.
J. Harper (1985)
Prevention of unwanted proteolysis during extraction of proteins from protease-rich tissue.
M. Hassel (1996)
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C. X. Cai (2019)