title: AAK1 identified as an inhibitor of neuregulin-1/ErbB4-dependent neurotrophic factor signaling using integrative chemical genomics and proteomics.
abstract: Target identification remains challenging for the field of chemical biology. We describe an integrative chemical genomic and proteomic approach combining the use of differentially active analogs of small molecule probes with stable isotope labeling by amino acids in cell culture-mediated affinity enrichment, followed by subsequent testing of candidate targets using RNA interference-mediated gene silencing. We applied this approach to characterizing the natural product K252a and its ability to potentiate neuregulin-1 (Nrg1)/ErbB4 (v-erb-a erythroblastic leukemia viral oncogene homolog 4)-dependent neurotrophic factor signaling and neuritogenesis. We show that AAK1 (adaptor-associated kinase 1) is a relevant target of K252a, and that the loss of AAK1 alters ErbB4 trafficking and expression levels, providing evidence for a previously unrecognized role for AAK1 in Nrg1-mediated neurotrophic factor signaling. Similar strategies should lead to the discovery of novel targets for therapeutic development.
authors.affiliation: Stanley Center for Psychiatric Research, 7 Cambridge Center, Cambridge, MA 02142, USA.
authors.forename: Letian
authors.initials: L
authors.lastname: Kuai
authors.forename: Shao-En
authors.initials: SE
authors.lastname: Ong
authors.forename: Jon M
authors.initials: JM
authors.lastname: Madison
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.forename: Jeremy R
authors.initials: JR
authors.lastname: Duvall
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Lewis
authors.forename: Catherine J
authors.initials: CJ
authors.lastname: Luce
authors.forename: Sean D
authors.initials: SD
authors.lastname: Conner
authors.forename: David A
authors.initials: DA
authors.lastname: Pearlman
authors.forename: John L
authors.initials: JL
authors.lastname: Wood
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Schreiber
authors.forename: Steven A
authors.initials: SA
authors.lastname: Carr
authors.forename: Edward M
authors.initials: EM
authors.lastname: Scolnick
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Haggarty
mesh_terms: D000818:Animals; D002227:Carbazoles / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D023281:Genomics / Q000379:methods; D006801:Humans; D026121:Indole Alkaloids / Q000378:metabolism; D008958:Models, Molecular; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D020890:Neuregulin-1 / Q000235:genetics / Q000378:metabolism*; D016501:Neurites / Q000378:metabolism; D016716:PC12 Cells; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D051381:Rats; D066247:Receptor, ErbB-4; D015398:Signal Transduction
chemical_list: D002227:Carbazoles; D026121:Indole Alkaloids; D009414:Nerve Growth Factors; D020890:Neuregulin-1; C049985:staurosporine aglycone; C587689:ERBB4 protein, human; D066246:ErbB Receptors; C587691:Erbb4 protein, rat; D066247:Receptor, ErbB-4; C562295:AAK1 protein, rat; D017346:Protein Serine-Threonine Kinases