Luminous proteins include primary light producers, such as aequorin, and secondary photoproteins that in some organisms red-shift light emission for better penetration in space. When expressed in heterologous systems, both types of proteins may act as versatile reporters capable of monitoring phenomena as diverse as calcium homoeostasis, protein sorting, gene expression, and so on. The Ca2+-sensitive photoprotein aequorin was targeted to defined intracellular locations (organelles, such as mitochondria, endoplasmic reticulum, sarcoplasmic reticulum, Golgi apparatus and nucleus, and cytoplasmic regions, such as the bulk cytosol and the subplasmalemmal rim), and was used to analyse Ca2+ homoeostasis at the subcellular level. We will discuss this application, reviewing its advantages and disadvantages and the experimental procedure. The applications of green fluorescent protein (GFP) are even broader. Indeed, the ability to molecularly engineer and recombinantly express a strongly fluorescent probe has provided a powerful tool for investigating a wide variety of biological events in live cells (e.g. tracking of endogenous proteins, labelling of intracellular structures, analysing promoter activity etc.). More recently, the demonstration that, using appropriate mutants and/or fusion proteins, GFP fluorescence can become sensitive to physiological parameters or activities (ion concentration, protease activity, etc.) has further expanded its applications and made GFP the favourite probe of cell biologists. We will here present two applications in the field of cell signalling, i.e. the use of GFP chimaeras for studying the recruitment of protein kinase C isoforms and the activity of intracellular proteases.
Skip Nav Destination
Article navigation
April 2001
-
Cover Image
Cover Image
- PDF Icon PDF LinkTable of Contents
Review Article|
February 26 2001
Recombinant aequorin and green fluorescent protein as valuable tools in the study of cell signalling
Anna CHIESA;
Anna CHIESA
1
*University of Ferrara, Department of Experimental and Diagnostic Medicine, Section of General Pathology, Via L. Borsari 46, 44100 Ferrara, Italy
Search for other works by this author on:
Elena RAPIZZI;
Elena RAPIZZI
1
*University of Ferrara, Department of Experimental and Diagnostic Medicine, Section of General Pathology, Via L. Borsari 46, 44100 Ferrara, Italy
Search for other works by this author on:
Valeria TOSELLO;
Valeria TOSELLO
*University of Ferrara, Department of Experimental and Diagnostic Medicine, Section of General Pathology, Via L. Borsari 46, 44100 Ferrara, Italy
Search for other works by this author on:
Paolo PINTON;
Paolo PINTON
†University of Padua, Department of Biomedical Sciences, Via G. Colombo 3, 35121 Padua, Italy
Search for other works by this author on:
Maddalena DE VIRGILIO;
Maddalena DE VIRGILIO
*University of Ferrara, Department of Experimental and Diagnostic Medicine, Section of General Pathology, Via L. Borsari 46, 44100 Ferrara, Italy
Search for other works by this author on:
Kevin E. FOGARTY;
Kevin E. FOGARTY
‡Biomedical Imaging Group, University of Massachusetts Medical School, 373 Plantation St., Worcester, MA 01655, U.S.A.
Search for other works by this author on:
Rosario RIZZUTO
Rosario RIZZUTO
2
*University of Ferrara, Department of Experimental and Diagnostic Medicine, Section of General Pathology, Via L. Borsari 46, 44100 Ferrara, Italy
2To whom correspondence should be addressed (e-mail rzr@dns.unife.it).
Search for other works by this author on:
Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2001
2001
Biochem J (2001) 355 (1): 1–12.
Citation
Anna CHIESA, Elena RAPIZZI, Valeria TOSELLO, Paolo PINTON, Maddalena DE VIRGILIO, Kevin E. FOGARTY, Rosario RIZZUTO; Recombinant aequorin and green fluorescent protein as valuable tools in the study of cell signalling. Biochem J 1 April 2001; 355 (1): 1–12. doi: https://doi.org/10.1042/bj3550001
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.