Assay for three-way interaction of protein phosphatase-1 (Glc7) with regulatory subunits plus phosphatase inhibitor-2.
A method is described using yeast conjugation to assay the interactions of a protein phosphatase-1 (PP1) inhibitor protein with holoenzymes formed in situ by expression of regulatory subunit fusion proteins that recruit endogenous Glc7, the yeast ortholog of PP1. Mutations in the canonical recognition motif VxF used to bind PP1 (Glc7) allow for analysis of direct from indirect (three-way) interactions.
Eto
Masumi
M
Department of Physiology, Thomas Jefferson College of Medicine, Philadelphia, PA, USA.
Brautigan
David L
DL
D001665:Binding Sites; D020134:Catalytic Domain; D020035:Holoenzymes / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D054645:Protein Phosphatase 1; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D000097847:Protein Phosphatase Inhibitory Proteins
D020035:Holoenzymes; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1; D011506:Proteins; C024833:protein phosphatase inhibitor-2; D000097847:Protein Phosphatase Inhibitory Proteins