title: Construction and analyses of elastically coupled multiple-motor systems.
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.
authors.affiliation: Departments of Chemistry and Bioengineering, Rice University, Houston, Texas, USA.
authors.forename: Arthur
authors.initials: A
authors.lastname: Rogers
authors.affiliation: Departments of Chemistry and Bioengineering, Rice University, Houston, Texas, USA.
authors.forename: Pamela E
authors.initials: PE
authors.lastname: Constantinou
authors.affiliation: Departments of Chemistry and Bioengineering, Rice University, Houston, Texas, USA.
authors.forename: D Kenneth
authors.initials: DK
authors.lastname: Jamison
authors.affiliation: Departments of Chemistry and Bioengineering, Rice University, Houston, Texas, USA.
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Driver
authors.affiliation: Departments of Chemistry and Bioengineering, Rice University, Houston, Texas, USA. Electronic address: diehl@rice.edu.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Diehl
mesh_terms: D001692:Biological Transport; D001696:Biomechanical Phenomena; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004548:Elasticity; D020409:Molecular Motor Proteins / Q000737:chemistry* / Q000378:metabolism; D052898:Optical Tweezers; D011108:Polymers / Q000737:chemistry* / Q000378:metabolism
chemical_list: D020409:Molecular Motor Proteins; D011108:Polymers; D004247:DNA
keywords: Cooperativity; Intracellular transport; Optical trapping; Self-assembly