Selective adhesion of functional microtubules to patterned silane surfaces

Abstract

We show that microtubule polymers can be immobilized selectively on lithographically patterned silane surfaces while retaining their native properties. Silane films were chemisorbed on polished silicon wafers or glass coverslips and patterned using a deep UV lithographic process developed at the Naval Research Laboratory. Hydrocarbon and fluorocarbon alkyl silanes, as well as amino and thiol terminal alkyl silanes, were investigated as substrates for microtubule adhesion with retention of biological activity. Microtubules were found to adhere strongly to amine terminal silanes while retaining the ability to act as substrates for the molecular motor protein kinesin. Aminosilane patterns with linewidths varying from 1 to 50 microns were produced lithographically and used to produce patterns of selectively adhered microtubules. Microtubules were partially aligned on the patterned lines by performing the immobilization in a fluid flow field. Patterns were imaged with atomic force microscopy and differential interference contrast microscopy. Motility assays were carried out using kinesin-coated beads and observed with differential interference contrast microscopy. Kinesin bead movement on the patterned microtubules was comparable to movement on microtubule control surfaces.

Dynamic photo-control of kinesin on a photoisomerizable monolayer – hydrolysis rate of ATP and motility of microtubules depending on the terminal group

“…One is that the photoisomerization induced a change in the interaction between the monolayer surface having positively charged amino groups and microtubules being rich in (1) (2) (4) (3) the negatively charged carboxylate groups. 52 The other one is that the photoresponsive monolayer affects to the activity of kinesin.…”

Protein Linear Molecular Motor‐Powered Nanodevices

“…Indeed, the first motility assays used naturally selfassembled actin filaments in the algae Nitella [72] and later artificially self-assembled F-actin paracrystals on lipid surfaces. [95] Early [4,30] and later work [90] demonstrated that microtubules adhere strongly to amine-terminated surfaces while retaining the ability to act as substrates for kinesin-coated beads. The critical issue of microtubule deactivation has also been addressed through stabilization with taxol [96] or glutaraldehyde.…”

“…Nonetheless, an early study [30] demonstrated confinement of the movement of kinesin-coated beads on microtubules that had been pre-aligned in a flow field and immobilized on aminosilane patterns. More recently, microfabricated electrodes and flow fields were used to transversally align filaments.…”

Active self-organization of microtubules in an inert chamber system

“…Currently, the in vitro motility assay is also used to determine the feasibility of constructing motor protein-based artificial biomachines, for which actin-myosin or MT-kinesin systems are considered promising building blocks. [8][9][10][11][12][13][14][15][16] New classes of devices including nanoscale molecular shuttles, 17,18 surface-imaging processes, 19 force measurements 20 and lab-on-a-chip devices 21 have been developed using knowledge obtained through the in vitro motility assay. To improve the organizational hierarchy of motor protein-based systems with emergent functions similar to those observed in natural systems, several active self-organization (AcSO) techniques have been developed.…”