English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Targeting of green fluorescent protein to neurodendocrine secretory granules: a new tool for real time studies of regulated protein secretion

Kaether, C., Salm, T., Glombrik, M., Almers, W., & Gerdes, H.-H. (1997). Targeting of green fluorescent protein to neurodendocrine secretory granules: a new tool for real time studies of regulated protein secretion. European Journal of Cell Biology: EJCB, 74(2), 133-142. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/9352218.

Item is

Basic

show hide
Genre: Journal Article
Alternative Title : Targeting of green fluorescent protein to neurodendocrine secretory granules: a new tool for real time studies of regulated protein secretion

Files

show Files
hide Files
:
EurJCellBiol_74_1997_133.pdf (Any fulltext), 17MB
 
File Permalink:
-
Name:
EurJCellBiol_74_1997_133.pdf
Description:
-
OA-Status:
Visibility:
Restricted (Max Planck Institute for Medical Research, MHMF; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Kaether, Christoph, Author
Salm, Thorsten, Author
Glombrik, Michael, Author
Almers, Wolfhard1, Author           
Gerdes, Hans-Herman, Author
Affiliations:
1Department of Molecular Cell Research, Max Planck Institute for Medical Research, Max Planck Society, ou_1497703              

Content

show
hide
Free keywords: GFP - chromogranin B - secretory granules - regulated secretion - real time
 Abstract: Human chromogranin B (hCgB), a soluble marker protein of neuroendocrine secretory granules, was fused to green fluorescent protein (GFP). Two GFP-mutants with different folding properties, S65T and EGFP, were used to produce two recombinant proteins, hCgB-GFP(S65T) and hCgB-EGFP, respectively. After transient expression only hCgB-EGFP elicited green fluorescence in the neuroendocrine cell line PC12. Pulse-chase experiments with [35S]sulfate followed by subcellular fractionation showed that hCgB-EGFP was sorted with high efficiency to immature secretory granules (ISG). Confocal microscopy revealed that fluorescent hCgB-EGFP colocalized largely with synaptotagmin, a membrane marker of secretory granules and synaptic-like microvesicles, and significantly with endogenous rat chromogranin B (rCgB), a soluble marker of secretory granules. Upon stimulation of transfected cells with 5 mM Ba2+ or by depolarization with 50 mM K+ hCgB-EGFP underwent regulated exocytosis. The dynamics of green fluorescent secretory granules beneath the plasma membrane (PM) of living PC12 cells were visualized by confocal microscopy. The majority of these vesicles did not move within 8.5 sec as if they were docked. In contrast, in NGF-induced cells most of the secretory granules beneath the somatic PM moved within the same time period whereas only little movement was observed in the neurites. These findings indicate that in differentiated PC12 cells the majority of the docking zones are not in the soma but are distributed along the neurites. In conclusion, the fusion protein hCgB-EGFP provides a powerful tool to study in real time vesicular traffic in the regulated pathway of protein secretion

Details

show
hide
Language(s): eng - English
 Dates: 1997-05-051997-06-051997-10
 Publication Status: Issued
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 665935
Other: 5601
URI: http://www.ncbi.nlm.nih.gov/pubmed/9352218
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: European Journal of Cell Biology : EJCB
  Other : Eur. J. Cell Biol.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Stuttgart : Wissenschaftliche Verlagsgesellschaft.
Pages: - Volume / Issue: 74 (2) Sequence Number: - Start / End Page: 133 - 142 Identifier: ISSN: 0070-2463
CoNE: https://pure.mpg.de/cone/journals/resource/954925486755