Work

Phosphatidylinositol 4,5-bisphosphate drives Ca2+-independent membrane penetration by the tandem C2 domain proteins synaptotagmin-1 and Doc2β

Year: 2019

Type: article

Abstract: Exocytosis mediates the release of neurotransmitters and hormones from neurons and neuroendocrine cells.Tandem C2 domain proteins in the synaptotagmin (syt) and double C2 domain (Doc2) families regulate exocytotic membrane fusion via direct interactions with Ca 2؉ and phospholipid bilayers.Syt1 is a fast-acting, low-affinity Ca 2؉ sensor that penetrates membranes upon binding Ca 2؉ to trigger synchronous vesicle fusion.The closely related Doc2␤ is a slow-acting, high-affinity Ca 2؉ sensor that triggers spontaneous and asynchronous vesicle fusion, but whether it also penetrates membranes is unknown.Both syt1 and Doc2␤ bind the dynamically regulated plasma membrane lipid phosphatidylinositol 4,5-bisphosphate (PIP 2 ), but it is unclear whether PIP 2 serves only as a membrane contact or enables specialized membrane-binding modes by these Ca 2؉ sensors.Furthermore, it has been shown that PIP 2 uncaging can trigger rapid, syt1-dependent exocytosis in the absence of Ca 2؉ influx, suggesting that current models for the action of these Ca 2؉ sensors are incomplete.Here, using a series of steady-state and time-resolved fluorescence measurements, we show that Doc2␤, like syt1, penetrates membranes in a Ca 2؉ -dependent manner.Unexpectedly, we observed that PIP 2 can drive membrane penetration by both syt1 and Doc2␤ in the absence of Ca 2؉ , providing a plausible mechanism for Ca 2؉ -independent, PIP 2 -dependent exocytosis.Quantitative measurements of penetration depth revealed that, in the presence of Ca 2؉ , PIP 2 drives Doc2␤, but not syt1, substantially deeper into the membrane, defining a biophysical regulatory mechanism specific to this high-affinity Ca 2؉ sensor.Our results provide evidence of a novel role for PIP 2 in regulating, and under some circumstances triggering, exocytosis.

Source: Journal of Biological Chemistry

DOI: 10.1074/jbc.ra119.007929

Authors: Mazdak M. Bradberry, Huan Bao, Xiaochu Lou, Edwin R. Chapman

Institutions: Howard Hughes Medical Institute, University of Wisconsin–Madison

Language: English

Topic: Cellular transport and secretion

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $3,430