Optimized localization analysis for single-molecule tracking and super-resolution microscopy.
AuthorsMortensen KI, Churchman LS, Spudich JA, Flyvbjerg H,
JournalNat Methods
PubMed ID20364147
'We optimally localized isolated fluorescent beads and molecules imaged as diffraction-limited spots, determined the orientation of molecules and present reliable formulas for the precision of various localization methods. Both theory and experimental data showed that unweighted least-squares fitting of a Gaussian squanders one-third of the available information, a popular formula ... More
ADP-induced rocking of the kinesin motor domain revealed by single-molecule fluorescence polarization microscopy.
AuthorsSosa H, Peterman EJ, Moerner WE, Goldstein LS
JournalNat Struct Biol
PubMed ID11373624
Kinesin is an ATP-driven molecular motor protein that moves processively along microtubules. Despite considerable research, the detailed mechanism of kinesin motion remains elusive. We applied an enhanced suite of single- and multiple-molecule fluorescence polarization microscopy assays to report the orientation and mobility of kinesin molecules bound to microtubules as a ... More
Nucleotide binding and hydrolysis induces a disorder-order transition in the kinesin neck-linker region.
AuthorsAsenjo AB, Weinberg Y, Sosa H
JournalNat Struct Mol Biol
PubMed ID16783374
Kinesin translocation is thought to occur by a conformational change in a region of the motor domain called the neck linker. However, most evidence supporting this hypothesis comes from monomeric constructs unable to move processively. To address this issue, we investigated the neck-linker configuration on microtubule-bound monomeric and dimeric kinesin ... More
Polarized fluorescence microscopy of individual and many kinesin motors bound to axonemal microtubules.
AuthorsPeterman EJ, Sosa H, Goldstein LS, Moerner WE
JournalBiophys J
PubMed ID11606296
Kinesin is a molecular motor that interacts with microtubules and uses the energy of ATP hydrolysis to produce force and movement in cells. To investigate the conformational changes associated with this mechanochemical energy conversion, we developed a fluorescence polarization microscope that allows us to obtain information on the orientation of ... More