Using the Lac repressor system to identify interacting proteins.
The present protocol describes a series of procedures to identify peptides interacting with PDZ domains. It is conceivable that the procedures can be applied to other purified protein modules or intact proteins without substantial modifications. With the deduced consensus combined with sequence information, it is possible to identify proteins present in the database with compatible sequences. If the expression of target protein and potential interacting candidate overlap temporally and spatially, biochemical and molecular experiments can be designed to study their physical and functional interactions.
Stricker
N L
NL
Department of Neuroscience, School of Medicine, Johns Hopkins University, Baltimore, Maryland 21205, USA.
Schatz
P
P
Li
M
M
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary; D019992:Databases as Topic; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D015723:Gene Library; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D056968:Lac Repressors; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D052248:Nitric Oxide Synthase Type I; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D018909:src Homology Domains
D001426:Bacterial Proteins; D018076:DNA, Complementary; D029968:Escherichia coli Proteins; D056968:Lac Repressors; C539064:LacI protein, E coli; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D005982:Glutathione Transferase