ORIGINAL ARTICLES  CELLULAR AND MOLECULAR ADVANCES IN THE STUDY OF INFLAMMATION 

Minerva Biotecnologica 2004 June;16(2):119-26

Copyright © 2004 EDIZIONI MINERVA MEDICA

language: English

Ca2+ homeostasis of intracellular compartments: measurements using the jellyfish photo-protein aequorin

Pinton P. ^{1, 2}, Bianchi K. ^{1, 2}, Prandini A. ^{1, 2}, Rimessi A. ^{1, 2}, Rizzuto R. ^{1, 2}

¹ Section of General Pathology, Department of Experimental and Diagnostic Medicine Telethon Center for Cell Imaging (TCCI), University of Ferrara, Ferrara, Italy; ² Interdisciplinary Center for the Study of Inflammation (ICSI) University of Ferrara, Ferrara, Italy

The devel­op­ment of molec­u­lar biol­o­gy tech­niques (which ­enable the mod­ifi­ca­tion and expres­sion of exog­e­nous cDNA in het­er­ol­o­gous ­cell ­types), has ­been respon­sible in ­recent ­years for the wide­spread use of pro­tein ­probes by ­cell biol­o­gists, for the meas­ure­ment of bio­log­i­cal param­e­ters. To ­this end, 2 ­main ­types of pro­teins are ­used, iso­lat­ed ­from a ­wide varie­ty of lumi­nes­cent organ­isms. The ­first ­group con­sists of chem­ilu­mi­nes­cent pro­teins. These are pro­teins ­that ­emit ­light, ­often asso­ciat­ed to phys­io­log­i­cal param­e­ters of inter­est, ­such as chang­es in ATP or Ca2+ con­cen­tra­tion. Since mam­mal­ian ­cells do not pos­sess endog­e­nous lumiu­nes­cent mole­cules, the use of ­these pro­teins is nor­mal­ly asso­ciat­ed to an excel­lent sig­nal-to-­noise ­ratio, as exem­pli­fied by aequ­or­in, a pho­to­pro­tein iso­lat­ed ­from Aequorea vic­tor­ia. The sec­ond ­group con­sists of flu­o­res­cent pro­teins. Among ­these, Aequorea ­victoria’s ­green flu­o­res­cent pro­tein (GFP) has attract­ed ­much atten­tion, ­since its expres­sion (or ­that of ­fusion prod­ucts ­with pro­teins of inter­est) gen­er­ates an ­intense sig­nal, ­which ­enables com­plex phe­nom­e­na (­such as orga­nelle struc­ture and ­their dynam­ic chang­es, pro­tein tar­get­ing etc.) to be fol­lowed in ­real ­time in liv­ing ­cells. This ­review ­presents ­some of the ­results ­obtained ­using aequ­or­in for stud­y­ing intra­cel­lu­lar Ca2+ homeo­sta­sis.