A peptide tag system for facile purification and single-molecule immobilization.
A peptide fusion tag and accompanying recombinant capture reagents have been developed on the basis of the peptide-PDZ domain interaction and affinity clamps, a new class of affinity reagent. This system allows for single-step purification under mild conditions and stable capture of a tagged protein. The subnanomolar affinity, high force resistance (>30 pN), small size ( approximately 25 kDa, approximately one-sixth of the size of IgG), and monomeric nature of the affinity clamp are all superior features for many applications, in particular single-molecule measurements.
Huang
Jin
J
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois 60637, USA.
Nagy
Stanislav S
SS
Koide
Akiko
A
Rock
Ronald S
RS
Koide
Shohei
S
D000345:Affinity Labels; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D006801:Humans; D007202:Indicators and Reagents; D008856:Microscopy, Fluorescence; D009218:Myosins / Q000737:chemistry* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry
D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D001710:Biotin; D019809:Streptavidin; D009218:Myosins