The mechanical role of VASP in an Arp2/3-complex-based motility assay

Sandy Suei; Rajveer Seyan; Philippe Noguera; John Manzi; Julie Plastino; Laurent Kreplak

doi:10.1016/j.jmb.2011.08.054

The mechanical role of VASP in an Arp2/3-complex-based motility assay

Sandy Suei, Rajveer Seyan, Philippe Noguera, John Manzi, Julie Plastino, Laurent Kreplak

Medicine, Science

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

The comet motility assay, inspired by Listeria locomotion, has been used extensively as an in vitro model to study the structural and motile properties of the actin cytoskeleton. However, there are no quantitative measurements of the mechanical properties of these actin comets. In this work, we use nanoindentation based on atomic force microscopy to measure the elastic modulus of actin comets grown on 1-μm-diameter beads in an Arp2/3 (actin-related proteins 2 and 3)-complex-dependent fashion in the absence and in the presence of VASP (vasodilator-stimulated phosphoprotein). Recruitment of VASP to the bead surface had no effect on the initial velocity or morphology of the comets. Instead, we observed an improved contact of the comets with the beads and an increased elastic modulus of the comets. The VASP-mediated increase in elastic modulus was dependent on both concentration and ionic strength. In conclusion, we propose that VASP plays a mechanical role in Arp2/3-complex-dependent motility by amplifying the elastic modulus of the thus assembled actin network and, consequently, by strengthening its cohesion for persistent protrusion.

Original language	English
Pages (from-to)	573-583
Number of pages	11
Journal	Journal of Molecular Biology
Volume	413
Issue number	3
DOIs	https://doi.org/10.1016/j.jmb.2011.08.054
Publication status	Published - Oct 28 2011

Bibliographical note

Funding Information:
The authors thank Dorothy Schafer for providing capping protein and Gijsje Koenderink and Corinne Gullekson for helpful discussions. S.S. acknowledges support from the Human Frontiers Science Program through a Young investigator grant awarded to J.P., L.K., and Gijsje Koenderink acknowledges support from the Natural Sciences and Engineering Research Council and the Canada Foundation for Innovation.

ASJC Scopus Subject Areas

Structural Biology
Molecular Biology

Access to Document

10.1016/j.jmb.2011.08.054

Cite this

@article{e9d6b66b9e5646f3b7e9fadeed686898,

title = "The mechanical role of VASP in an Arp2/3-complex-based motility assay",

abstract = "The comet motility assay, inspired by Listeria locomotion, has been used extensively as an in vitro model to study the structural and motile properties of the actin cytoskeleton. However, there are no quantitative measurements of the mechanical properties of these actin comets. In this work, we use nanoindentation based on atomic force microscopy to measure the elastic modulus of actin comets grown on 1-μm-diameter beads in an Arp2/3 (actin-related proteins 2 and 3)-complex-dependent fashion in the absence and in the presence of VASP (vasodilator-stimulated phosphoprotein). Recruitment of VASP to the bead surface had no effect on the initial velocity or morphology of the comets. Instead, we observed an improved contact of the comets with the beads and an increased elastic modulus of the comets. The VASP-mediated increase in elastic modulus was dependent on both concentration and ionic strength. In conclusion, we propose that VASP plays a mechanical role in Arp2/3-complex-dependent motility by amplifying the elastic modulus of the thus assembled actin network and, consequently, by strengthening its cohesion for persistent protrusion.",

author = "Sandy Suei and Rajveer Seyan and Philippe Noguera and John Manzi and Julie Plastino and Laurent Kreplak",

note = "Funding Information: The authors thank Dorothy Schafer for providing capping protein and Gijsje Koenderink and Corinne Gullekson for helpful discussions. S.S. acknowledges support from the Human Frontiers Science Program through a Young investigator grant awarded to J.P., L.K., and Gijsje Koenderink acknowledges support from the Natural Sciences and Engineering Research Council and the Canada Foundation for Innovation.",

year = "2011",

month = oct,

day = "28",

doi = "10.1016/j.jmb.2011.08.054",

language = "English",

volume = "413",

pages = "573--583",

journal = "Journal of Molecular Biology",

issn = "0022-2836",

publisher = "Academic Press Inc.",

number = "3",

}

TY - JOUR

T1 - The mechanical role of VASP in an Arp2/3-complex-based motility assay

AU - Suei, Sandy

AU - Seyan, Rajveer

AU - Noguera, Philippe

AU - Manzi, John

AU - Plastino, Julie

AU - Kreplak, Laurent

N1 - Funding Information: The authors thank Dorothy Schafer for providing capping protein and Gijsje Koenderink and Corinne Gullekson for helpful discussions. S.S. acknowledges support from the Human Frontiers Science Program through a Young investigator grant awarded to J.P., L.K., and Gijsje Koenderink acknowledges support from the Natural Sciences and Engineering Research Council and the Canada Foundation for Innovation.

PY - 2011/10/28

Y1 - 2011/10/28

N2 - The comet motility assay, inspired by Listeria locomotion, has been used extensively as an in vitro model to study the structural and motile properties of the actin cytoskeleton. However, there are no quantitative measurements of the mechanical properties of these actin comets. In this work, we use nanoindentation based on atomic force microscopy to measure the elastic modulus of actin comets grown on 1-μm-diameter beads in an Arp2/3 (actin-related proteins 2 and 3)-complex-dependent fashion in the absence and in the presence of VASP (vasodilator-stimulated phosphoprotein). Recruitment of VASP to the bead surface had no effect on the initial velocity or morphology of the comets. Instead, we observed an improved contact of the comets with the beads and an increased elastic modulus of the comets. The VASP-mediated increase in elastic modulus was dependent on both concentration and ionic strength. In conclusion, we propose that VASP plays a mechanical role in Arp2/3-complex-dependent motility by amplifying the elastic modulus of the thus assembled actin network and, consequently, by strengthening its cohesion for persistent protrusion.

AB - The comet motility assay, inspired by Listeria locomotion, has been used extensively as an in vitro model to study the structural and motile properties of the actin cytoskeleton. However, there are no quantitative measurements of the mechanical properties of these actin comets. In this work, we use nanoindentation based on atomic force microscopy to measure the elastic modulus of actin comets grown on 1-μm-diameter beads in an Arp2/3 (actin-related proteins 2 and 3)-complex-dependent fashion in the absence and in the presence of VASP (vasodilator-stimulated phosphoprotein). Recruitment of VASP to the bead surface had no effect on the initial velocity or morphology of the comets. Instead, we observed an improved contact of the comets with the beads and an increased elastic modulus of the comets. The VASP-mediated increase in elastic modulus was dependent on both concentration and ionic strength. In conclusion, we propose that VASP plays a mechanical role in Arp2/3-complex-dependent motility by amplifying the elastic modulus of the thus assembled actin network and, consequently, by strengthening its cohesion for persistent protrusion.

UR - http://www.scopus.com/inward/record.url?scp=80054705187&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80054705187&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2011.08.054

DO - 10.1016/j.jmb.2011.08.054

M3 - Article

C2 - 21925510

AN - SCOPUS:80054705187

SN - 0022-2836

VL - 413

SP - 573

EP - 583

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 3

ER -

The mechanical role of VASP in an Arp2/3-complex-based motility assay

Abstract

Bibliographical note

ASJC Scopus Subject Areas

Access to Document

Other files and links

Fingerprint

Cite this