Hundreds of myosin 10s are pushed to the tips of filopodia and could cause traffic jams on actin.
actin
biochemistry
cell biology
chemical biology
filopodia
human
myosin 10
protein quantitation
Journal
eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614
Informations de publication
Date de publication:
31 Oct 2024
31 Oct 2024
Historique:
medline:
1
11
2024
pubmed:
1
11
2024
entrez:
31
10
2024
Statut:
epublish
Résumé
Myosin 10 (Myo10) is a motor protein known for its role in filopodia formation. Although Myo10-driven filopodial dynamics have been characterized, there is no information about the absolute number of Myo10 molecules during the filopodial lifecycle. To better understand molecular stoichiometries and packing restraints in filopodia, we measured Myo10 abundance in these structures. We combined SDS-PAGE densitometry with epifluorescence microscopy to quantitate HaloTag-labeled Myo10 in U2OS cells. About 6% of total intracellular Myo10 localizes to filopodia, where it enriches at opposite cellular ends. Hundreds of Myo10s are in a typical filopodium, and their distribution across filopodia is log-normal. Some filopodial tips even contain more Myo10 than accessible binding sites on the actin filament bundle. Live-cell movies reveal a dense cluster of over a hundred Myo10 molecules that initiates filopodial elongation. Hundreds of Myo10 molecules continue to accumulate during filopodial growth, but accumulation ceases when retraction begins. Rates of filopodial elongation, second-phase elongation, and retraction are inversely related to Myo10 quantities. Our estimates of Myo10 molecules in filopodia provide insight into the physics of packing Myo10, its cargo, and other filopodia-associated proteins in narrow membrane compartments. Our protocol provides a framework for future work analyzing Myo10 abundance and distribution upon perturbation.
Identifiants
pubmed: 39480891
doi: 10.7554/eLife.90603
pii: 90603
doi:
pii:
Substances chimiques
Myosins
EC 3.6.4.1
MYO10 protein, human
0
Actins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIGMS NIH HHS
ID : T32 GM144292
Pays : United States
Organisme : National Science Foundation
ID : 2140001
Organisme : NIGMS NIH HHS
ID : R01 GM124272
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM149073
Pays : United States
Commentaires et corrections
Type : UpdateOf
Informations de copyright
© 2023, Shangguan and Rock.
Déclaration de conflit d'intérêts
JS No competing interests declared, RR RSR is a consultant for Cyntegron Therapeutics
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