Next Article in Journal
Development and Validation of an Ultrasensitive Procalcitonin Sandwich Immunoassay
Previous Article in Journal
A Computational Workflow Translates a 58-Gene Signature to a Formalin-Fixed, Paraffin-Embedded Sample-Based Companion Diagnostic for Personalized Treatment of the BRAF-Mutation-Like Subtype of Colorectal Cancers
Article Menu

Export Article

Open AccessArticle
High-Throughput 2017, 6(4), 17; doi:10.3390/ht6040017

Application of High-Throughput Assays to Examine Phospho-Modulation of the Late Steps of Regulated Exocytosis

1
Department of Molecular Physiology, and the WSU Molecular Medicine Research Group, School of Medicine, Western Sydney University, Campbelltown, NSW 2560, Australia
2
Faculty of Applied Health Sciences and Faculty of Mathematics and Science, Brock University, St. Catharines, ON L2S 3A1, Canada
Present address: Department of Pharmacology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
*
Author to whom correspondence should be addressed.
Academic Editor: Massimo Negrini
Received: 20 September 2017 / Revised: 1 November 2017 / Accepted: 6 November 2017 / Published: 13 November 2017
View Full-Text   |   Download PDF [2338 KB, uploaded 13 November 2017]   |  

Abstract

Abstract: Regulated exocytosis enables a range of physiological functions including neurotransmission, and the late steps (i.e., docking, priming and Ca2+-triggered membrane fusion) are modulated by a highly conserved set of proteins and lipids. Many of the molecular components and biochemical interactions required have been identified; the precise mechanistic steps they modulate and the biochemical interactions that need to occur across steps are still the subject of intense investigation. Particularly, although the involvement of phosphorylation in modulating exocytosis has been intensively investigated over the past three decades, it is unclear which phosphorylation events are a conserved part of the fundamental fusion mechanism and/or serve as part of the physiological fusion machine (e.g., to modulate Ca2+ sensitivity). Here, the homotypic fusion of cortical vesicles was monitored by utilizing new high-throughput, cost-effective assays to assess the influence of 17 small molecule phospho-modulators on docking/priming, Ca2+ sensitivity and membrane fusion. Specific phosphatases and casein kinase 2 are implicated in modulating the Ca2+ sensitivity of fusion, whereas sphingosine kinase is implicated in modulating the ability of vesicles to fuse. These results indicate the presence of multiple kinases and phosphatases on the vesicles and critical phosphorylation sites on vesicle membrane proteins and lipids that directly influence late steps of regulated exocytosis. View Full-Text
Keywords: phosphorylation; docking; priming; membrane fusion; calcium sensitivity; lipids; secretory vesicles phosphorylation; docking; priming; membrane fusion; calcium sensitivity; lipids; secretory vesicles
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Abbineni, P.S.; Coorssen, J.R. Application of High-Throughput Assays to Examine Phospho-Modulation of the Late Steps of Regulated Exocytosis. High-Throughput 2017, 6, 17.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
High-Throughput EISSN 2571-5135 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top