Tethering forces of secretory granules measured with optical tweezers.
Fusion of a vesicle with its target membrane is preceded by tethering or docking. However, the physical mechanism of vesicle-tethering is unknown. To study this mechanism, we used eosinophil secretory granules, which undergo stimulated homotypic fusion events inside the cell during degranulation. Using a dual optical trap system, we observed tether formation between isolated eosinophil secretory granules. The results show that secretory granules interact stochastically with a target membrane forming physical tethers linking the vesicle and target membrane, rather than via interactions with the cytoskeleton. The necessary components are membrane-associated, and the addition of cytosolic components is not required. Tether-lifetime measurements as a function of applied mechanical force revealed at least three kinetically distinct tethered states. The tethered-state lifetimes of isolated eosinophil granules match the residence times of chromaffin granules at the plasma membrane in intact cells, suggesting that the tethering mechanisms reported here may represent the physiological mechanisms of vesicle-tethering in the cell.
Valero
Vicente
V
School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.
Nevian
Thomas
T
Ho
Dominik
D
Lindau
Manfred
M
D000818:Animals; D002462:Cell Membrane / Q000502:physiology* / Q000648:ultrastructure; D002478:Cells, Cultured; D004548:Elasticity; D004804:Eosinophils / Q000502:physiology* / Q000648:ultrastructure; D006736:Horses; D052898:Optical Tweezers; D022142:Secretory Vesicles / Q000502:physiology* / Q000648:ultrastructure; D013314:Stress, Mechanical