Mapping protease substrates by using a biotinylated phage substrate library. 2006

Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
Combinatorial Biology Unit, Biosciences Division, Argonne National Laboratory, Argonne, IL 60439, USA.

We describe a bacteriophage M13 substrate library encoding the AviTag (BirA substrate) and combinatorial heptamer peptides displayed at the N terminus of the mature form of capsid protein III. Phages are biotinylated efficiently (> or = 50%) when grown in E. coli cells coexpressing BirA, and such viral particles can be immobilized on a streptavidin-coated support and released by protease cleavage within the combinatorial peptide. We have used this library to map the specificity of human Factor Xa and a neuropeptidase, neurolysin (EC3.4.24.16). Validation by analysis of isolated peptide substrates has revealed that neurolysin recognizes the motif hydrophobic-X-Pro-Arg-hydrophobic, where Arg-hydrophobic is the scissile bond.

UI MeSH Term Description Entries
D008666 Metalloendopeptidases ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism. Metallo-Endoproteinases,Metalloendopeptidase
D010447 Peptide Hydrolases Hydrolases that specifically cleave the peptide bonds found in PROTEINS and PEPTIDES. Examples of sub-subclasses for this group include EXOPEPTIDASES and ENDOPEPTIDASES. Peptidase,Peptidases,Peptide Hydrolase,Protease,Proteases,Proteinase,Proteinases,Proteolytic Enzyme,Proteolytic Enzymes,Esteroproteases,Enzyme, Proteolytic,Hydrolase, Peptide
D010455 Peptides Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are considered to be larger versions of peptides that can form into complex structures such as ENZYMES and RECEPTORS. Peptide,Polypeptide,Polypeptides
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D013058 Mass Spectrometry An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers. Mass Spectroscopy,Spectrometry, Mass,Spectroscopy, Mass,Spectrum Analysis, Mass,Analysis, Mass Spectrum,Mass Spectrum Analysis,Analyses, Mass Spectrum,Mass Spectrum Analyses,Spectrum Analyses, Mass
D015951 Factor Xa Activated form of factor X that participates in both the intrinsic and extrinsic pathways of blood coagulation. It catalyzes the conversion of prothrombin to thrombin in conjunction with other cofactors. Autoprothrombin C,Coagulation Factor Xa,Factor X, Activated,Thrombokinase,Activated Factor X,Blood Coagulation Factor X, Activated,Factor 10A,Factor Ten A,Factor Xa, Coagulation
D017104 Bacteriophage M13 Temperate bacteriophage of the genus INOVIRUS which infects enterobacteria, especially E. coli. It is a filamentous phage consisting of single-stranded DNA and is circularly permuted. Bacteriophage fd,Coliphage M13,Coliphage fd,Enterobacteria phage M13,Enterobacteria phage fd,M13 Phage,Phage M13,Phage fd,fd Phage,M13 Phages,M13, Coliphage,Phage, M13,Phage, fd,Phages, M13,Phages, fd,fd Phages
D019151 Peptide Library A collection of cloned peptides, or chemically synthesized peptides, frequently consisting of all possible combinations of amino acids making up an n-amino acid peptide. Phage Display Peptide Library,Random Peptide Library,Peptide Phage Display Library,Phage Display Library, Peptide,Synthetic Peptide Combinatorial Library,Synthetic Peptide Library,Libraries, Peptide,Libraries, Random Peptide,Libraries, Synthetic Peptide,Library, Peptide,Library, Random Peptide,Library, Synthetic Peptide,Peptide Libraries,Peptide Libraries, Random,Peptide Libraries, Synthetic,Peptide Library, Random,Peptide Library, Synthetic,Random Peptide Libraries,Synthetic Peptide Libraries
D019921 Biotinylation Incorporation of biotinyl groups into molecules. Biotinylations

Related Publications

Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
May 1993, Science (New York, N.Y.),
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
January 2013, PloS one,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
August 2016, Cell chemical biology,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
September 2000, Virology,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
December 2012, Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
March 2005, Combinatorial chemistry & high throughput screening,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
September 1997, Virology,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
January 2004, The Journal of biological chemistry,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
February 1994, Analytical biochemistry,
Michael D Scholle, and Ushma Kriplani, and Amanda Pabon, and Kamakshi Sishtla, and Marc J Glucksman, and Brian K Kay
July 2020, Chembiochem : a European journal of chemical biology,
Copied contents to your clipboard!