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A Journal on Biotechnology and Molecular Biology

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Minerva Biotecnologica 2001 December;13(4):281-5

language: English

Use of pep­tide micro­ar­rays for assay­ing phos­pho-depen­dent pro­tein inter­ac­tions

Violina S. 1, Francioso F. 1, Venturoli L. 1, Tosi L. 2, Marastoni M. 3, Carinci P. 2, Carella M. 4, Stanziale P. 5, Evangelisti R. 1

­1 Università degli Studi di Ferrara, Laboratorio di Genomica Funzionale, Sezione di Istologia, Dipartimento di Morfologia ed Embriologia;
2 Università degli Studi di Bologna, Centro Ricerca Fondazione CARISBO, Istituto di Istologia ed Embriologia;
3 Università degli Studi di Ferrara, Dipartimento di Scienze Farmaceutiche;
4 TIGEM, Napoli;
5 IRCCS Casa di Sollievo della Sofferenza, S. Giovanni Rotondo, Foggia, Italy


Back­ground. ­High through­put pro­tein ­assays ­need to be ­devised to pro­cess and val­i­date ­data pro­duced by the cur­rent genom­ics pro­jects. ­While DNA micro­ar­rays are now wide­ly ­used in main­stream sci­en­tif­ic experi­ments, pro­tein ­chips are ­still in the devel­op­ing ­phase.
Meth­ods. We eval­u­at­ed dif­fer­ent con­di­tions of pep­tide spot­ting for pro­duc­ing pep­tide micro­ar­rays. Our over­all ­aims are ­those of ­using micro­ar­ray tech­nol­o­gy to ­assess and val­i­date pro­tein-pro­tein inter­ac­tions. ­Here we ­show ­that pep­tide micro­ar­rays can be ­used in ­order to ­assay phos­phod­e­pen­dent and ace­tyl ­lysine depen­dent inter­ac­tions. ­High affin­ity gener­ic con­text or spe­cif­ic anti­bod­ies ­were ­used. Dif­fer­ent meth­ods of label­ing and detec­tions ­have ­been test­ed.
­Results. Opti­mal ­results ­were gen­er­at­ed by ­using cova­lent link­ing to acti­vat­ed ­glass ­slides and ordi­nary cya­nines.
Con­clu­sions. We are in the pro­cess of test­ing ­weak pro­tein-pep­tide inter­ac­tions by ­using ­these ­same pep­tide micro­ar­rays, and of trans­pos­ing a sim­i­lar tech­nique to the ­study of ­enzyme sub­strates.

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