Non-polymerizable monomeric actin: Difference between revisions
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=== | ===Nucleotide-dependent structural changes of AP-actin=== | ||
<table width='450' align='left' cellpadding='5'><tr><td rowspan='2'> </td><td bgcolor='#d0d0d0'><applet load=Image:2HF3 2HF4 10state morph4.pdb' size='450' align='right' scene='User:Thomas_E_Sladewski/Sandbox_1/10state_morph_scene2/2' /></td></tr><tr><td bgcolor='#d0d0d0'>Morph of the [[lac repressor]] complexed with DNA showing the differences between non-specific binding (straight DNA) vs. specific recognition of the operator sequence (kinked DNA). Whether the binding kinks the DNA, or simply stabilizes a pre-existing kink, is unknown. [[Lac repressor#Specific Binding|Details]].</td></tr></table> | <table width='450' align='left' cellpadding='5'><tr><td rowspan='2'> </td><td bgcolor='#d0d0d0'><applet load=Image:2HF3 2HF4 10state morph4.pdb' size='450' align='right' scene='User:Thomas_E_Sladewski/Sandbox_1/10state_morph_scene2/2' /></td></tr><tr><td bgcolor='#d0d0d0'>Morph of the [[lac repressor]] complexed with DNA showing the differences between non-specific binding (straight DNA) vs. specific recognition of the operator sequence (kinked DNA). Whether the binding kinks the DNA, or simply stabilizes a pre-existing kink, is unknown. [[Lac repressor#Specific Binding|Details]].</td></tr></table> | ||
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Revision as of 06:36, 12 April 2011
Non-polymerizable monomeric actin or AP-actin is an Sf9-expressed cytoplasmic actin harboring two point mutations that prevent the monomer from polymerizing into actin filaments. These mutations allow for the crystallization of actin without the use of specific toxins or actin-binding proteins that may influence the structure. The crystal structure of AP-actin has been solved for the ADP-bound form (2HF3) and the ATP-bound form (2HF4). These two structures are shown below as a morph between the two states.
Structural features of actinStructural features of actin
The actin monomer, or , contains four structural domains: (residues 1-32, 70-144 and 138-375), (residues 33-69), (residues 145-180 and 270-337) and (residues 181-269). These domains can be classified as largely alpha+beta connected by loops which are shown in some structural analysis to undergo significant nucleotide dependent structural changes. The is located in domain 2 (residues 40-51). This loop is not shown in the crystal structure of AP-actin because it was found to be disordered in both the ATP and ADP-bound state, conflicting with other reports. This is discussed in more detail below. The or sensor loop is located in domain 1 (residues 70-78) and is though to contain important residues for sensing the nucleotide state. The (residues 165-172) is located in domain 3 and is important for the binding of WH2 domain-containing proteins. The (residues 11-16) and the (residues 154-161) are contained in the nucleotide-binding cleft.
AP-actin is a cytoplasmic actin encoded from the Drosophila 5C actin gene. It shares 98.7% sequence homology with human γ-cytoplasmic actin. AP-actin contains two point mutations, that render it incapable of polymerizing into actin filaments. These mutations were shown to have little effect on ATP hydrolysis or long range structural effects.
Nucleotide-dependent structural changes of AP-actinNucleotide-dependent structural changes of AP-actin
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Morph of the lac repressor complexed with DNA showing the differences between non-specific binding (straight DNA) vs. specific recognition of the operator sequence (kinked DNA). Whether the binding kinks the DNA, or simply stabilizes a pre-existing kink, is unknown. Details. |
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MechanismMechanism
About this StructureAbout this Structure
This is where I will add my text. 2hf4 is a 1 chain structure of Actin with sequence from Drosophila melanogaster. Full crystallographic information is available from OCA.
See AlsoSee Also
ReferenceReference
- ↑ Rould MA, Wan Q, Joel PB, Lowey S, Trybus KM. Crystal structures of expressed non-polymerizable monomeric actin in the ADP and ATP states. J Biol Chem. 2006 Oct 20;281(42):31909-19. Epub 2006 Aug 18. PMID:16920713 doi:M601973200