Fig. 2.

Characterization of the putative CaM/CML substrates on the Arabidopsis protein microarrays. (A) Protein microarray containing 1,133 purified protein preparations arrayed in duplicate spots on FAST slides was probed with anti-MYC antibodies. An enlarged image of one block is shown to the right of the protein microarray. (B) Representative targets of CaM/CML proteins. The top row shows the amount of each protein preparation on the microarray as detected with anti-MYC antibodies. The bottom row (negative control) shows the signal detected on the microarray for the same protein preparations after probing with the AtCaM1 in the presence of the Ca²⁺ chelator EGTA. Lanes: 1, At2g23080; 2, At1g54610; 3, At5g20810; 4, At4g23650; 5, At1g74740. (C) Representation of the total number of CaM/CML targets identified on the protein microarrays. Estimated numbers of substrates binding to each CaM/CML protein are shown above each column. (D) Representation of CaM/CML target specificity. Estimated numbers of substrates predicted to bind a number of CaM/CML are shown above each column. (E) Hierarchical clustering analysis of predicted CaM/CML-interacting proteins. Columns represent various CaMs/CMLs, and rows represent CaM-binding proteins (see SI Table 8 for gene names in rows). The resultant cladogram, generated by single linkage hierarchical clustering, is shown at the top. Red depicts interaction, and black indicates no interaction. An enlarged high-resolution image is shown as SI Fig. 5.