Format

Send to

Choose Destination
Methods Enzymol. 2014;540:189-204. doi: 10.1016/B978-0-12-397924-7.00011-X.

Construction and analyses of elastically coupled multiple-motor systems.

Author information

1
Departments of Chemistry and Bioengineering, Rice University, Houston, Texas, USA.
2
Departments of Chemistry and Bioengineering, Rice University, Houston, Texas, USA. Electronic address: diehl@rice.edu.

Abstract

Precision analyses of the collective motor behaviors have become important to dissecting mechanisms underlying the trafficking of subcellular commodities in eukaryotic cells. Here, we describe a synthetic approach to create structurally defined multiple protein complexes containing two elastically coupled motor molecules. Motors are connected using a simple DNA-scaffolding molecule and DNA-conjugated, artificial protein polymers that function as tunable elastic linkers. The procedure to self-assemble these components produces complexes in high synthetic yield and allows individual multiple-motor systems to be interrogated at the single-complex level. Methods to evaluate cooperative motor responses in a static optical trap are also discussed. While enabling the average transport properties of single/noninteracting and coupled motors to be compared, these procedures can provide insight into the extent to which motors cooperate productively via load sharing as well as the roles loading-rate-dependent phenomena play in collective motor functions.

KEYWORDS:

Cooperativity; Intracellular transport; Optical trapping; Self-assembly

[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center