Green fluorescent protein labeling of cytoskeletal structures--novel targeting approach based on leucine zippers.
Green fluorescent protein (GFP) is a valuable marker for intracellular protein localization. However the fusion of GFP with structural proteins can alter their properties, resulting in a loss of fusion protein localization, decreased GFP fluorescence or both. We describe a novel targeting approach based on noncovalent heterodimerization of GFP and cytoplasmic structural proteins. The formation of structural protein/GFP complexes was mediated by modified leucine zipper protein spacers designed to form high-affinity heterodimers. The complexes localized accurately to specific sites within cells, providing selective fluorescence labeling of subcellular structures such as microfilaments or focal contacts.
Katz
B Z
BZ
National Institute of Dental Research, NIH, Bethesda, MD, USA.
Krylov
D
D
Aota
S
S
Olive
M
M
Vinson
C
C
Yamada
K M
KM
D000199:Actins / Q000235:genetics; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D005347:Fibroblasts; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D016350:Leucine Zippers / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016596:Vinculin / Q000378:metabolism
D000199:Actins; D003598:Cytoskeletal Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D016596:Vinculin; D049452:Green Fluorescent Proteins