title: MASPIT: three-hybrid trap for quantitative proteome fingerprinting of small molecule-protein interactions in mammalian cells.
abstract: Organic small molecules generally act by perturbing the function of one or more cellular target proteins, the identification of which is essential to an understanding of the molecular basis of drug action. Here we describe the application of methotrexate-linked small molecule ligands to a mammalian three-hybrid interaction trap for proteome-wide identification of small molecule targets, quantification of the targeting potency of unmodified small molecules for such targets in intact cells, and screening for inhibitors of small molecule-protein interactions. During the course of this study we also identified the pyrido[2,3-d]pyrimidine PD173955, a known SRC kinase inhibitor, as a potent inhibitor of several ephrin receptor tyrosine kinases. This finding could perhaps be exploited in the design of inhibitors for this kinase subfamily, members of which have been implicated in the pathogenesis of various diseases, including cancer.
authors.affiliation: GPC Biotech Incorporated, Waltham, Massachusetts 02451, USA.
authors.forename: Maureen
authors.initials: M
authors.lastname: Caligiuri
authors.forename: Lisa
authors.initials: L
authors.lastname: Molz
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.forename: Faith
authors.initials: F
authors.lastname: Kaplan
authors.forename: Jimmy P
authors.initials: JP
authors.lastname: Xu
authors.forename: Jiangwen Z
authors.initials: JZ
authors.lastname: Majeti
authors.forename: Rebeca
authors.initials: R
authors.lastname: Ramos-Kelsey
authors.forename: Krishna
authors.initials: K
authors.lastname: Murthi
authors.forename: Sam
authors.initials: S
authors.lastname: Lievens
authors.forename: Jan
authors.initials: J
authors.lastname: Tavernier
authors.forename: Nikolai
authors.initials: N
authors.lastname: Kley
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D018076:DNA, Complementary; D005434:Flow Cytometry; D006801:Humans; D007700:Kinetics; D011506:Proteins / Q000737:chemistry*; D020543:Proteome; D011728:Pyridones / Q000737:chemistry / Q000494:pharmacology; D011743:Pyrimidines / Q000737:chemistry / Q000494:pharmacology
chemical_list: D018076:DNA, Complementary; C403095:PD 173955; D011506:Proteins; D020543:Proteome; D011728:Pyridones; D011743:Pyrimidines