title: Rational targeting of active-site tyrosine residues using sulfonyl fluoride probes.
abstract: This work describes the first rational targeting of tyrosine residues in a protein binding site by small-molecule covalent probes. Specific tyrosine residues in the active site of the mRNA-decapping scavenger enzyme DcpS were modified using reactive sulfonyl fluoride covalent inhibitors. Structure-based molecular design was used to create an alkyne-tagged probe bearing the sulfonyl fluoride warhead, thus enabling the efficient capture of the protein from a complex proteome. Use of the probe in competition experiments with a diaminoquinazoline DcpS inhibitor permitted the quantification of intracellular target occupancy. As a result, diaminoquinazoline upregulators of survival motor neuron protein that are used for the treatment of spinal muscular atrophy were confirmed as inhibitors of DcpS in human primary cells. This work illustrates the utility of sulfonyl fluoride probes designed to react with specific tyrosine residues of a protein and augments the chemical biology toolkit by these probes uses in target validation and molecular pharmacology.
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Erik C
authors.initials: EC
authors.lastname: Hett
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Hua
authors.initials: H
authors.lastname: Xu
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Kieran F
authors.initials: KF
authors.lastname: Geoghegan
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Ariamala
authors.initials: A
authors.lastname: Gopalsamy
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Kyne
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Carol A
authors.initials: CA
authors.lastname: Menard
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Arjun
authors.initials: A
authors.lastname: Narayanan
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Mihir D
authors.initials: MD
authors.lastname: Parikh
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Shenping
authors.initials: S
authors.lastname: Liu
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Lee
authors.initials: L
authors.lastname: Roberts
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Ralph P
authors.initials: RP
authors.lastname: Robinson
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Tones
authors.affiliation: †Worldwide Medicinal Chemistry and ‡Rare Disease Research Unit, Pfizer, 610 Main Street, Cambridge, Massachusetts 02139, United States.|§Structural Biology and Biophysics, Worldwide Medicinal Chemistry, ∥Worldwide Medicinal Chemistry, and ⊥Primary Pharmacology Group, Pfizer, Eastern Point Road, Groton, Connecticut 06340, United States.
authors.forename: Lyn H
authors.initials: LH
authors.lastname: Jones
mesh_terms: D020134:Catalytic Domain; D002478:Cells, Cultured; D060326:Chemistry Techniques, Synthetic; D018360:Crystallography, X-Ray; D004722:Endoribonucleases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D058990:Molecular Targeted Therapy / Q000379:methods; D013329:Structure-Activity Relationship; D013441:Sulfinic Acids / Q000737:chemistry*; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D015335:Molecular Probes; D013441:Sulfinic Acids; D014443:Tyrosine; C048899:sulfuryl fluoride; D004722:Endoribonucleases; C485332:DcpS protein, human