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. 1997 Sep 16;94(19):10467–10472. doi: 10.1073/pnas.94.19.10467

Figure 4.

Figure 4

KAPP interactions with the CLV1 kinase domain. (A) GST-KI protein was labeled at the heart muscle kinase site with [γ-32P]ATP and incubated with a filter containing immobilized GST-CKD (lane 1), GST-CKD1-(lane 2), GST-CKDK720E (lane 3), GST-CKD treated with AP (lane 4), GST-CKD treated with AP in the presence of 100 mM PO4− (lane 5), and GST-CKD treated with AP, then allowed to autophosphorylate in the presence of MnCl2 and ATP (lane 6). (Upper) The Coomassie blue-stained gel showing the relative amounts of fusion protein in each lane. (Lower) The autoradiograph. (B) The GST-KAPP protein was incubated alone (lane 1) or with GST-CKD (lane 2), GST-CKD1-(lane 3), or GST-CKDK720E (lane 4) during an autophosphorylation reaction. Equal amounts of GST-CKD, GST-CKD1-1, and GST-CKDK720E were added in each reaction. (C) 32P-labeled GST-CKD was incubated with full-length GST-KAPP protein (lanes 1 and 2), or without GST-KAPP (lanes 3 and 4), and with either MgCl2/MnCl2 (lanes 1 and 3) or EDTA (lanes 2 and 4). Lane 5 is an untreated control. (Upper) Coomassie blue-stained gel showing the approximately equal loading of GST-CKD in each lane. (Lower) The autoradiogram.