Expression of scFv antibodies in Xenopus embryos to disrupt protein function: implications for large-scale evaluation of the embryonic proteome.
We evaluated the use of single-chain antibody (scFv) expression as a tool to disrupt the function of specific proteins in embryos of the frog, Xenopus laevis. The expression of scFvs that recognize the bone morphogenetic protein receptor (ALK3) or the fibroblast growth factor receptor1 (FGFR1) as endoplasmic reticulum-anchored proteins caused distinct developmental defects that were virtually indistinguishable from the defects caused by expression of the dominant negative forms of each receptor. These results demonstrate that scFvs from phage-display libraries can be readily fashioned into effective and specific inhibitors of signaling pathways in developing embryos. In addition, as several effective scFvs against a specific target can be isolated rapidly, this approach represents a valuable new tool for large-scale functional analysis of the embryonic proteome.
Abler
Lisa L
LL
Department of Biomolecular Chemistry, University of Wisconsin, Madison, Wisconsin 53706, USA.
Sheets
Michael D
MD
D030201:Activin Receptors, Type I / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D052005:Bone Morphogenetic Protein Receptors, Type I; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D019151:Peptide Library; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases; D017434:Protein Structure, Tertiary; D020543:Proteome / Q000032:analysis; D012333:RNA, Messenger / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism / Q000502:physiology*; D051496:Receptor, Fibroblast Growth Factor, Type 1; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism / Q000502:physiology*; D017978:Receptors, Growth Factor; D015398:Signal Transduction; D013997:Time Factors; D014982:Xenopus laevis / Q000196:embryology*
D000906:Antibodies; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D020543:Proteome; D012333:RNA, Messenger; D017468:Receptors, Fibroblast Growth Factor; D017978:Receptors, Growth Factor; D020794:Receptor Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; D052005:Bone Morphogenetic Protein Receptors, Type I