ABA-regulated SNRK2 Protein Kinase: Difference between revisions

| <applet load='3ujg' size='400' frame='true' align='left' caption='3uc4 - Apo SnRK2.6' scene = '55/559985/Aposnrk2_6/6' /><br  clear='both'>'''3uc4 scenes''' <Br><scene name='55/559985/Aposnrk2_6/7'>1. Default scene</scene> The catalytic domain of SnRK2.6 is typical of [[Eukaryotic Protein Kinase Catalytic Domain]]) except for an additional α-helix (shown as strands) in the small lobe, which is formed by SNRK2 box sequence.  <Br><scene name='55/559985/Aposnrk2_6critical/3'>2. Important structures:</scene> The activation segment (with unresolved gap), including the D of the DFG motif in ball and stick, is blue. The catalytic loop, including the D of the DLKLEN motif in ball and stick, is orchid. Subdomain III, including its invariant E in ball and stick, is gold. The invariant K of subdomain II is in chartreuse. The SnRK2 box is turquoise. The C-terminal domain, that includes the ABA box is unresolved. The arrangement of the residues in ball and stick around the active site, indicate that this structure is in a partially active state in spite of its unphosphorylated activation loop. This is possibly due to the interaction of the SNRK2 box helix with subdomain III.<ref name = "Ng2011"/>. The interaction between these helices is similar to the interaction of helices in the complex between <scene name='55/559985/Cdk2-cyclin/3'>cyclin-dependent protein kinase 2 (CDK2) and cyclin</scene> [[1w98]].  Here we see that subdomain III of the protein kinase (opaque blue) is stablized by interaction with a helix from cyclin (opaque gold). The positioning of subdomain III by this interaction is critical for for formation of the active site.<ref>PMID:15660127</ref>. <br><br><br><br><br><br><br><br><br><br>

| <applet load='3ujg' size='400' frame='true' align='left' caption='3ujg - SnRK2.6-HAB1' scene = '55/559985/Aposnrk2_6/2' /><Br clear='both'>'''3ujg scenes'''<Br><scene name='55/559985/Aposnrk2_6/2'>1. Default Scene</scene> The two enzymes are bound via interface their active sites. The phosphatase inactivates the kinase by dephosphorylating the kinase activation loop and by sterically blocking the kinase active site. The complex was constructed as a fusion protein with a 6His-tag at the N-terminus of SnRK2.6 (residues 11–362) fused to HAB1(172–511) via a GSGSAGSAAGS linker. Mutations of D296A and E297A in SnRK2.6 were introduced at the crystal packing interface to reduce surface entropy. <br><scene name='55/559985/Ost1hab1_critical/3'>2. Important structures in SnRK2.6</scene> The same structures as in the left scene are shown. The fully resolved activation segment extends into the phosphatase's active site and is unphosphorylated. Residues 319-362 of SnRK2.6, which includes the ABA box, and the GSGSAGSAAGS linker are not resolved. The disorganization of the residues shown in ball and stick, with most pointing away from the active site, indicates that the catalytic domain is in the inactive state.<Br><scene name='55/559985/Ost1hab1_interaction/2'>3. Zone of interaction</scene> The activation loop (blue trace) of SnRK2.6 is inserted into the catalytic site (marked by the magnesium ions) of the phosphatase. The phosphorylatable residue of the activation loop S175 (CPK ball and stick) is positioned near the magnesium ions. W385 of the phosphatase (brown ball and stick) in turn protrudes into the kinase's active site, where it interacts with residues R139 and Glu144 (CPK ball and stick) of the catalytic loop (orchid trace) and I183 of the activation loop.<br><scene name='55/559985/Tetherbinding/1'> 5. Proposed interaction zone </scene> SnRK2.6 is shown in blue cartoon, and HAB1 in gold spacefill. The ABA box sequence is not resolved, but it would extend from the C-terminal end of the SNRK2 box helix (cyan helix). It is proposed that the ABA box sequence, which is highly acidic, binds to a patch of basic residues (blue) on the surface of the phosphatase.<ref name= "Soon2012"/>