Application of a chimeric green fluorescent protein to study protein-protein interactions.
The green fluorescent protein (GFP) of the jellyfish Aequorea victoria is an emerging tool to monitor gene expression in situ and in vivo. Because of its fluorescence properties, when GFP is fused in-frame to a specific protein of interest, various aspects of the behavior of this protein can be analyzed noninvasively. Here we describe a fusion between GFP and human calmodulin-like protein (CLP) and show that this protein retains fluorescence and known characteristics of CLP, including Ca(2+)-dependent interaction with phenyl-Sepharose and interaction with a specific cellular target protein. The results suggest a novel application for GFP fusion proteins in the rapid, nonradioactive detection of interacting proteins on gel overlays.
Garamszegi
N
N
Mayo Graduate School, Mayo Clinic/Foundation, Rochester, MN 55905, USA.
Garamszegi
Z P
ZP
Rogers
M S
MS
DeMarco
S J
SJ
Strehler
E E
EE
D019921:Biotinylation; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D015342:DNA Probes; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D013698:Templates, Genetic
D002135:Calcium-Binding Proteins; D017931:DNA Primers; D015342:DNA Probes; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D015247:Deoxyribonuclease HindIII; D015252:Deoxyribonucleases, Type II Site-Specific; C029312:GGTACC-specific type II deoxyribonucleases; D002118:Calcium