Covalent attachment of actin filaments to Tween 80 coated polystyrene beads for cargo transportation

In this manuscript, a new strategy has been reported for circumscribed covalent attachment of barbed and pointed ends of actin filaments to polystyrene beads. A comparative study of attachment of actin filaments to polystyrene beads was performed by blocking functionally active sites on polystyrene beads with nonionic detergents such as Tween 20, Tween 80 and polyethylene glycol (PEG). Effective blocking of active sites was obtained with Tween 80 at 0.1% concentration. Attachment of single bundle of actin filament to bead was assessed by rotational motion of bead tailed actin in front and lateral view. Velocity of actin filaments attached to different size of beads in in-vitro motility assay was calculated to ascertain their attachments. Velocity of actin attached to 1.0 and 3.0 microm polystyrene beads was reduced to 3.0-4.0 and 0.0-1.0 microm/s, respectively as compared to free actin velocity of 4.0-6.0 microm/s. Single point attachment of actin filaments to different size of beads was assessed by decrease in sliding velocity. Present study provides insight into the actin-myosin based molecular motor systems for drug delivery and biosensors applications.

[1]  Ronald D. Vale,et al.  The case for a common ancestor: kinesin and myosin motor proteins and G proteins , 1998, Journal of Muscle Research & Cell Motility.

[2]  J. Spudich,et al.  Assays for actin sliding movement over myosin-coated surfaces. , 1991, Methods in enzymology.

[3]  G. Langford Actin- and microtubule-dependent organelle motors: interrelationships between the two motility systems. , 1995, Current opinion in cell biology.

[4]  R. P. Bajpai,et al.  Covalent immobilization of myosin forin-vitro motility of actin , 2005 .

[5]  S. Ishiwata,et al.  Preparation of bead-tailed actin filaments: estimation of the torque produced by the sliding force in an in vitro motility assay. , 1996, Biophysical journal.

[6]  Ronald D Vale,et al.  The Molecular Motor Toolbox for Intracellular Transport , 2003, Cell.

[7]  Lars Montelius,et al.  Nanotechnology and actomyosin motility in vitro on different surface chemistries , 2004 .

[8]  S. Brown,et al.  Nucleation of polar actin filament assembly by a positively charged surface , 1979, The Journal of cell biology.

[9]  L. Montelius,et al.  Actin-Based Molecular Motors for Cargo Transportation in Nanotechnology— Potentials and Challenges , 2005, IEEE Transactions on Advanced Packaging.

[10]  John M. Walker,et al.  Molecular Motors , 2007, Methods in Molecular Biology™.

[11]  Manfred Schliwa,et al.  Molecular motors , 2003, Nature.

[12]  S. Ishiwata,et al.  Stepwise motion of an actin filament over a small number of heavy meromyosin molecules is revealed in an in vitro motility assay. , 1994, Journal of biochemistry.