Figure 2: AHP1 is S-nitrosylated at Cys 115.
From: S-nitrosylation of phosphotransfer proteins represses cytokinin signaling
(a) S-nitrosylation of recombinant His-tagged AHPs (SNO) in vitro. (b,c) S-nitrosylation of endogenous AHP1 (b) and transgenic AHP1-GFP (c) proteins in Columbia-0 (Col-0) and AHP1::AHP1-GFP seedlings. (d) Accumulation of S-nitrosylated AHP1-GFP in wild-type (WT) and gsnor1-3 plants. A quantitative analysis of the data is shown at the right (n=3). Error bars indicate s.d.; **indicates P<0.01 (Student’s t-test). (e) Mass spectrometric analysis of the tryptic fragments of GSNO-treated GST4CS-AHP1 recombinant protein. The N-terminal fragments (b2 through b5) are charged with biotin, thereby identifying Cys 115 as the S-nitrosylated residue. (f) S-nitrosylation of GST4CS-AHP1 and GST4CS-AHP1C115S recombinant proteins. (g) S-nitrosylation of AHP1-GFP and AHP1C115S-GFP proteins in vivo.
