Using peptide libraries to identify optimal cleavage motifs for proteolytic enzymes.
Proteases and peptidases are involved in a vast array of fundamental cellular processes, including cell growth, survival, motility, death, and differentiation, and can be important players in multicellular systems such as angiogenesis, inflammation, and immunity. Though long considered to be essentially digestive enzymes that mediate complete degradation of their substrates, many proteases are now known to be highly site specific. Knowledge of the cleavage site motif for a protease or peptidase can be useful in the design of substrates and inhibitors for the enzyme, and can also provide insight into its biological function through the identification and characterization of its protein substrates. Here, we describe in detail methodology that allows for the rapid and general determination of optimal recognition sequences for proteolytic enzymes.
Turk
Benjamin E
BE
Division of Signal Transduction, Beth Israel Deaconess Medical Center, Department of Cell Biology, Harvard Medical School, 330 Brookline Avenue, Boston, MA 02215, USA.
Cantley
Lewis C
LC
D000107:Acetylation; D001360:Avidin / Q000737:chemistry; D001665:Binding Sites / Q000235:genetics; D019921:Biotinylation; D005456:Fluorescent Dyes / Q000737:chemistry; D010447:Peptide Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000235:genetics / Q000378:metabolism; D011480:Protease Inhibitors / Q000138:chemical synthesis / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity
D005456:Fluorescent Dyes; D019151:Peptide Library; D010455:Peptides; D011480:Protease Inhibitors; C066402:avidin-agarose; D001360:Avidin; D012685:Sepharose; D010447:Peptide Hydrolases