From Proteopedia

Revision as of 10:02, 26 March 2010

F-Actin

Filamentous actin (F-actin) is also referred to as microfilament ^[1] and is a highly conserved proteinous component found near ubiquitously in eukaryotic cytoskeletons. F-actin and other actin proteins generally provide a structural role to the cell.

Introduction

Actin is found in nearly all eukaryotic cells and is known primarily for its function as a structural and translocation protein. It also has an ATPase function, as it hydrolyzes ATP --> ADP + Pi and undergoes conformational changes with each hydrolysis. Actin belongs to the actin superfamily, which includes other proteins such as Hsp70 and hexokinase, because of its nucelotide-dependent conformational change^[2]. Prokaryotes are not known to have actin, but do however have an actin homologue, MreB^[3]

Actin occurs in two forms: globular actin (G-actin), the free monomeric units of actin, and filamentous actin (F-actin) which is the polymer form. These two forms exist in a dynamic equilibrium with one another as ATP-associated polymerization and depolymerization occur continuously within the cell.

Assembly

G-actin is the free monomeric form of actin which transitions to F-actin. The structures of globular and filamentous actin are distinct from one another in numerous ways, despite the fact that G-actin comprises F-actin. When the monomeric actin becomes polymerized into F-actin, the unit becomes flattened. G-actin appears to have more ion ligands in its structure, and also has the ligand RHO as opposed to 4-methyl histidine as found in the F-actin structure.

Formation of F-actin is a dynamic process of assembly and disassembly. The transition between G- and F-actin begins with a stabilized oligomer of actin units forms through a nucleation-condensation type fold pattern^[4]. Addition of monomeric units to either end subsequently occurs, however, because of a difference in charge polarity in the two ends, there is preferential addition to what is termed the "plus (+) end" or the "barbed-end". On the opposite end, the "minus (-) end" or the "pointed end", there is preferential dissociation of actin units^[5]. The rate of actin association is ten-fold greater than that of dissociation, meaning that f-actin moves toward growth in length^[6].

Structure

History of the structure

The F-actin protein was discovered by Straub in 1942. The structure was speculated based on a low-resolution x-ray crystallograph found in 1990 by Holmes et al. and over this time, despite the importance of F-actin in eukaryotic cells, this speculated structure was accepted. A higher resolution structure was only recently deposited in the PDB databank in Decemeber 2008 by Oda et al. ^[7].

Polymer F-actin

F-actin has the appearance of two right-handed helices. It is actually composed of repeats of 13 actin units for every 6 left-handed turns, spanning a length of 350 Å. ^[8]. Including the ADP and Ca²⁺, the F-actin molecule as shown here consists of 377 residues (43kDa), two major domains separated by a nucleotide-binding cleft^[9]. Depending on the state of the bound nucleotide, the most stable conformation of F-actin changes. In its ATP and ADP + Pi nucleotide bound states, it has a closed binding cleft. In its ADP only bound state, it has a wider binding cleft^[10]. A characteristic trait of actin is that the domains remain twisted relative to one another, despite the nucleotide-state-dependent conformational changes^[11].

Domains

Domain movement is made possible by rotation about the shown in purple. These occur in an alpha-helix (Ile136-Gly146) sequence that does not belong to either domain^[12].

Stability

The flattened folded form of F-actin requires different stabilization mechanisms than the free monomeric G-actin form. Stability of the f-actin complex is achieved by a series of salt bridge formations involving polar and charged residues, including HIC73, a methylated histidine residue. Additional stability is believed to arise from cross-subunit interactions between Leu110 and Thr194.

Function

Structural Role

Enzymatic Role

Active Site

The cleavage of the gamma-phosphoryl from the bound ATP is a result of a conformational change upon binding that moves the Gln137 residue closer to the ATP-Ca2+ ligand. Release of the inorganic phosphate occurs via the conformational change of the flexible "D-loop" into an ordered alpha-helix^[13].

References

1. ↑ Microfilament - Wikipedia, the free encyclopedia. http://en.wikipedia.org/wiki/Microfilaments. Date accessed: March 16th, 2010.
2. ↑ Graceffa
3. ↑ Oda
4. ↑ pfaendtner
5. ↑ mitchinson
6. ↑ clasier
7. ↑ Oda T, Iwasa M, Aihara T, Maéda Y, and Narita A. 2009. The nature of the globular-to fibrous actin transition. Nature,457(7228):441-445. PMID: 19158791
8. ↑ Holmes, K.C., Popp, D., Gebhard, W. and Kabsch, W. 1990. Atomic model of the actin filament. Nature,347(6288):44-49. PMID: 2395461
9. ↑ oda
10. ↑ pfaendtner
11. ↑ oda
12. ↑ graceffa
13. ↑ graceffa