Nucleotide binding and hydrolysis induces a disorder-order transition in the kinesin neck-linker region

Ana B. Asenjo, Yonatan Weinberg, Hernando J. Sosa

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

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 constructs using single-molecule fluorescence polarization microscopy. We found that the neck-linker region (i) is very mobile in the absence of nucleotides and during steady walking, (ii) decreases mobility and aligns along the microtubule axis in the presence of AMPPNP or ADP+AlF-4, (iii) is mostly ordered in the monomeric constructs in the presence of ADP+AlF- 4, and (iv) is closer to parallel to the microtubule axis in the dimeric constructs. These results support the proposed role of the neck linker and suggest a coordination mechanism between the two motor domains in the dimer.

Original languageEnglish (US)
Pages (from-to)648-654
Number of pages7
JournalNature Structural and Molecular Biology
Volume13
Issue number7
DOIs
StatePublished - Jul 2006

Fingerprint

Kinesin
Hydrolysis
Neck
Nucleotides
Microtubules
Adenosine Diphosphate
Adenylyl Imidodiphosphate
Polarization Microscopy
Fluorescence Polarization
Fluorescence Microscopy
Walking

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

@article{d281c2af69644dcba3b3a4aea765d278,
title = "Nucleotide binding and hydrolysis induces a disorder-order transition in the kinesin neck-linker region",
abstract = "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 constructs using single-molecule fluorescence polarization microscopy. We found that the neck-linker region (i) is very mobile in the absence of nucleotides and during steady walking, (ii) decreases mobility and aligns along the microtubule axis in the presence of AMPPNP or ADP+AlF-4, (iii) is mostly ordered in the monomeric constructs in the presence of ADP+AlF- 4, and (iv) is closer to parallel to the microtubule axis in the dimeric constructs. These results support the proposed role of the neck linker and suggest a coordination mechanism between the two motor domains in the dimer.",
author = "Asenjo, {Ana B.} and Yonatan Weinberg and Sosa, {Hernando J.}",
year = "2006",
month = "7",
doi = "10.1038/nsmb1109",
language = "English (US)",
volume = "13",
pages = "648--654",
journal = "Nature Structural and Molecular Biology",
issn = "1545-9993",
publisher = "Nature Publishing Group",
number = "7",

}

TY - JOUR

T1 - Nucleotide binding and hydrolysis induces a disorder-order transition in the kinesin neck-linker region

AU - Asenjo, Ana B.

AU - Weinberg, Yonatan

AU - Sosa, Hernando J.

PY - 2006/7

Y1 - 2006/7

N2 - 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 constructs using single-molecule fluorescence polarization microscopy. We found that the neck-linker region (i) is very mobile in the absence of nucleotides and during steady walking, (ii) decreases mobility and aligns along the microtubule axis in the presence of AMPPNP or ADP+AlF-4, (iii) is mostly ordered in the monomeric constructs in the presence of ADP+AlF- 4, and (iv) is closer to parallel to the microtubule axis in the dimeric constructs. These results support the proposed role of the neck linker and suggest a coordination mechanism between the two motor domains in the dimer.

AB - 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 constructs using single-molecule fluorescence polarization microscopy. We found that the neck-linker region (i) is very mobile in the absence of nucleotides and during steady walking, (ii) decreases mobility and aligns along the microtubule axis in the presence of AMPPNP or ADP+AlF-4, (iii) is mostly ordered in the monomeric constructs in the presence of ADP+AlF- 4, and (iv) is closer to parallel to the microtubule axis in the dimeric constructs. These results support the proposed role of the neck linker and suggest a coordination mechanism between the two motor domains in the dimer.

UR - http://www.scopus.com/inward/record.url?scp=33745839864&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745839864&partnerID=8YFLogxK

U2 - 10.1038/nsmb1109

DO - 10.1038/nsmb1109

M3 - Article

C2 - 16783374

AN - SCOPUS:33745839864

VL - 13

SP - 648

EP - 654

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

SN - 1545-9993

IS - 7

ER -