Analysis of the trk NGF receptor tyrosine kinase using recombinant fusion proteins.
Nerve growth factor (NGF) represents a family of structurally related trophic factors, including brain-derived neurotrophin factor (BDNF), neurotrophin-3 (NT-3), NT-4, and NT-5. These neurotrophin factors interact with two classes of receptors, the trk receptor tyrosine kinase family, and the low affinity p75 neurotrophin receptor. To study potential ligand-receptor interactions, recombinant trk fusion proteins have been constructed, and pan-trk polyclonal antisera directed against the cytoplasmic tyrosine kinase domain have been generated. The recombinant proteins were assessed for in vitro kinase activity and for the ability of K-252a to inhibit phosphorylation. Antibodies made against the fusion protein recognize all trk family members, and are effective in immunoprecipitation of affinity-crosslinked receptors. Comparative crosslinking indicates that NGF can recognize all trk receptor members, illustrating the large number of potential ligand-receptor interactions between neurotrophins and their receptors.
Horvath
C M
CM
Department of Cell Biology and Anatomy, Cornell University Medical College, New York 10021.
Wolven
A
A
Machadeo
D
D
Huber
J
J
Boter
L
L
Benedetti
M
M
Hempstead
B
B
Chao
M V
MV
D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D002227:Carbazoles / Q000494:pharmacology; D003432:Cross-Linking Reagents; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D026121:Indole Alkaloids; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000378:metabolism; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020917:Receptor, trkA; D017475:Receptors, Nerve Growth Factor / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000906:Antibodies; D002227:Carbazoles; D003432:Cross-Linking Reagents; D018076:DNA, Complementary; D026121:Indole Alkaloids; D009414:Nerve Growth Factors; D011518:Proto-Oncogene Proteins; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; C049985:staurosporine aglycone; D020794:Receptor Protein-Tyrosine Kinases; D020917:Receptor, trkA