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.
National Institute of Dental Research, NIH, Bethesda, MD, USA.
B Z
BZ
Katz
D
D
Krylov
S
S
Aota
M
M
Olive
C
C
Vinson
K M
KM
Yamada
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