Quantitative analysis of self-association and mobility of annexin A4 at the plasma membrane.

Abstract:

:Annexins, found in most eukaryotic species, are cytosolic proteins that are able to bind negatively-charged phospholipids in a calcium-dependent manner. Annexin A4 (AnxA4) has been implicated in diverse cellular processes, including the regulation of exocytosis and ion-transport; however, its precise mechanistic role is not fully understood. AnxA4 has been shown to aggregate on lipid layers upon Ca(2+) binding in vitro, a characteristic that may be critical for its function. We have utilized advanced fluorescence microscopy to discern details on the mobility and self-assembly of AnxA4 after Ca(2+) influx at the plasma membrane in living cells. Total internal reflection microscopy in combination with Förster resonance energy transfer reveals that there is a delay between initial plasma membrane binding and the beginning of self-assembly and this process continues after the cytoplasmic pool has completely relocated. Number-and-brightness analysis suggests that the predominant membrane bound mobile form of the protein is trimeric. There also exists a pool of AnxA4 that forms highly immobile aggregates at the membrane. Fluorescence recovery after photobleaching suggests that the relative proportion of these two forms varies and is correlated with membrane morphology.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Crosby KC,Postma M,Hink MA,Zeelenberg CH,Adjobo-Hermans MJ,Gadella TW

doi

10.1016/j.bpj.2013.02.057

subject

Has Abstract

pub_date

2013-05-07 00:00:00

pages

1875-85

issue

9

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(13)00365-2

journal_volume

104

pub_type

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