000057862 001__ 57862
000057862 005__ 20180515100241.0
000057862 0247_ $$2doi$$a10.1371/journal.pone.0102572
000057862 0248_ $$2sideral$$a91898
000057862 037__ $$aART-2014-91898
000057862 041__ $$aeng
000057862 100__ $$aChakraborty, Sandeep
000057862 245__ $$aFRET based quantification and screening technology platform for the interactions of Leukocyte Function-associated Antigen-1 (LFA-1) with InterCellular Adhesion Molecule-1 (ICAM-1)
000057862 260__ $$c2014
000057862 5060_ $$aAccess copy available to the general public$$fUnrestricted
000057862 5203_ $$aThe interaction between leukocyte function-associated antigen-1(LFA-1) and intercellular adhesion molecule-1 (ICAM-1) plays a pivotal role in cellular adhesion including the extravasation and inflammatory response of leukocytes, and also in the formation of immunological synapse. However, irregular expressions of LFA-1 or ICAM-1 or both may lead to autoimmune diseases, metastasis cancer, etc. Thus, the LFA-1/ICAM-1 interaction may serve as a potential therapeutic target for the treatment of these diseases. Here, we developed one simple ‘in solution’ steady state fluorescence resonance energy transfer (FRET) technique to obtain the dissociation constant (Kd) of the interaction between LFA-1 and ICAM-1. Moreover, we developed the assay into a screening platform to identify peptides and small molecules that inhibit the LFA-1/ICAM-1 interaction. For the FRET pair, we used Alexa Fluor 488-LFA-1 conjugate as donor and Alexa Fluor 555-human recombinant ICAM-1 (D1-D2-Fc) as acceptor. From our quantitative FRET analysis, the Kd between LFA-1 and D1-D2-Fc was determined to be 17.93±1.34 nM. Both the Kd determination and screening assay were performed in a 96-well plate platform, providing the opportunity to develop it into a high-throughput assay. This is the first reported work which applies FRET based technique to determine Kd as well as classifying inhibitors of the LFA-1/ICAM-1 interaction.
000057862 536__ $$9info:eu-repo/grantAgreement/ES/MINECO-FEDER/SAF2011-25390
000057862 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000057862 590__ $$a3.234$$b2014
000057862 591__ $$aMULTIDISCIPLINARY SCIENCES$$b9 / 57 = 0.158$$c2014$$dQ1$$eT1
000057862 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000057862 700__ $$aNúñez, David
000057862 700__ $$aHu, Shih-Yang
000057862 700__ $$aDomingo, María Pilar
000057862 700__ $$0(orcid)0000-0003-0154-0730$$aPardo, Julian$$uUniversidad de Zaragoza
000057862 700__ $$aKarmenyan, Artashes
000057862 700__ $$aGálvez, Eva M
000057862 700__ $$aChiou, Arthur
000057862 7102_ $$11008$$2566$$aUniversidad de Zaragoza$$bDepartamento de Microbiología, Medicina Preventiva y Salud Pública$$cInmunología
000057862 773__ $$g9, 7 (2014), e102572 [13 pp]$$pPLoS One$$tPLoS One$$x1932-6203
000057862 8564_ $$s628023$$uhttp://zaguan.unizar.es/record/57862/files/texto_completo.pdf$$yVersión publicada
000057862 8564_ $$s122121$$uhttp://zaguan.unizar.es/record/57862/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000057862 909CO $$ooai:zaguan.unizar.es:57862$$particulos$$pdriver
000057862 951__ $$a2018-05-15-09:59:17
000057862 980__ $$aARTICLE