Targeting of synaptotagmin to neurite terminals in neuronally differentiated PC12 cells

Krasnov, P. A., Enikolopov, G. (April 2000) Targeting of synaptotagmin to neurite terminals in neuronally differentiated PC12 cells. Journal of Cell Science, 113 (8). pp. 1389-1404. ISSN 0021-9533

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URL: http://www.ncbi.nlm.nih.gov/pubmed/10725222

Abstract

We have investigated structural elements that determine the accumulation of synaptotagmin, a major synaptic vesicle protein, in neurite terminals of neuronally differentiated neuroendocrine pheochromocytoma PC12 cells. We performed extensive deletion and point mutagenesis of rat synaptotagmin II, expressed mutant proteins in PC12 cells differentiated by nerve growth factor (NGF) and monitored their intracellular distribution by immunofluorescence. We found a structural element located at the carboxy-terminal domain nf the synaptotagmin molecule, which is necessary for its accumulation at the terminal. Using alanine-scanning mutagenesis, we have identified two amino acids in this element, tryptophan W405 and leucine L408, that are critical for correct targeting of synaptotagmin II to neurite terminals. Changing either one of them to alanine prevents the accumulation of the protein at the terminals, These amino acids are evolutionarily conserved throughout the entire synaptotagmin family and also among synaptotagmin-related proteins, suggesting that different synaptotagmins may have similar mechanisms of targeting to neuronal cell terminals.

Item Type: Paper
Uncontrolled Keywords: synaptotagmin PC12 cell neurite terminal protein targeting ALPHA-LATROTOXIN RECEPTOR SYNAPTIC VESICLE PROTEIN DENSE-CORE VESICLES MEMBRANE-PROTEIN SECRETORY PATHWAY AXONAL-TRANSPORT PLASMA-MEMBRANE CALCIUM SENSOR COOH TERMINUS RAT-BRAIN
Subjects: organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > tissues types and functions > synaptogenesis
CSHL Authors:
Communities: CSHL labs > Enikopolov lab
Depositing User: Matt Covey
Date: April 2000
Date Deposited: 20 Dec 2013 15:45
Last Modified: 20 Dec 2013 15:45
Related URLs:
URI: https://repository.cshl.edu/id/eprint/29136

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