espectroscopia

ESPEKTROSKOPIA MINTEGIA

Bringing single-molecule spectroscopy to macromolecular
protein complexes.
Single-molecule protein studies (Bankako molekulen teknika proteinen azterketerako.)
Single-molecule techniques have become potent tools for the discovery of novel protein
mechanisms by allowing high spatiotemporal resolution. Sub-nanometer resolution, the
ultimate scale of biologicalsystems, has been reached with single-molecule fluorescence
microscopy[1] and spectroscopy[2]; single-molecule force and torque spectroscopy[3, 4];
atomic force microscopy[5] and spectroscopy[6]; and nanopores[7]. Measurements can be
carried out on the biologically relevant time scales of microseconds to minutes.
Banakako molekulen teknikek oso tresna garrantzitsu bilakatu egin da proteinenmekanismo berrientzako erresoluzio espazio-tenporal altua baimentzen. Erresoluzio sub-nanometrikoa, sistema biologikoetan azken eskala, garatu egin da banakako-molekulen fluoreszentzia mikroskopiarekin eta espektroskopiarekin; eta nanoporoekin. Neurketak biologikoki nabarmen den denbora eskaletan egin daiteke, mikrosegundotik minutuetara.
Among these techniques, single-molecule fluorescencespectroscopy has enabled researchers to unveil the action mechanism of a protein by imaging protein activity in real time. Benefitting from advances in general microscopy[1], detection devices[8], and fluorophore physics[9, 10], single-molecule fluorescence spectroscopy has reached sub-nanometer spatial resolution[11] and microsecond temporal resolution[12, 13]. Single-molecule fluorescence imaging iscarried out primarily with total internal reflection, confocal, and zero-mode waveguide microscopy[2]. Among these, total internal reflection fluorescence microscopy has been employed by several research teams in the development of novel techniques described in this review[14-17]. With total internal reflection microscopy, a localized fluorescence spot on a CCD (charge-coupled device) screen representstheinteraction of a fluorescent molecule with a surface-immobilized molecule. The time of thisdocking event and the ensuing time trajectory deliver valuable kinetic information on biochemical processes such as protein complex assemblies, protein-protein interactions, protein-nucleic acid interactions, and protein-lipid interactions.
Un punto fluorescente localizado en una pantalla CCD representala interaccion de una molecula fluorescente con una molecula immovilizada en la superficie. El tiempo de este evento de acoplamiento y la subsiguiente trayectoria de tiempo ofrecen información valiosa cinético en procesos bioquímicos como conjuntos complejos de proteína, interacciones proteína-proteína, interacciones de ácidos nucleicos de proteína y las interacciones proteína-lípidos.
Teknikahauen artean, banakako molekulen fluoreszentzia espektroskopia baimendu die ikertzaileei proteinen akzio mekanismoak ezagutaraztea imajinatzen nola diren proteinaren aktibitatea denbora errealean. Onura egiten dio oinarrizko espektroskopiari, detekzio sistemei, eta fluoroforoen fisikari, banakako molekulen fluoreszentzia espektroskopia subnanometroraino heldu da erresoluzio espazialean etamikrosegundoraino erresoluzio denboralean. Banakako molekulen fluoreszentziaren irudiak lortzen da nagusiki barne islapen totalarekin, konfokalarekin eta zero-era uhin gida mikroskopiarekin. Hauen artean, barne islapen totaleko fluoreszentzia mikroskopia erabili egin da ikerketa talde askotan txosten honetan agertzen diren teknika berrien garapenerako. Barne islapen totaleko mikroskopiarekin, CCDpantaila batean mugatuta agertzen den puntu fluoereszentea irudikatzen du molekula fluoreszente baten eta immobilizatuta dagoen molekulen elkarrekintzak. Akoplamenduaren denbora eta bere jarduera denboran prozesu biokimikoen informazio zinetikoa ematen digu, adibidez: Proteina konplexu multzoak, proteina-proteina elkarrekintzak, proteinen azido nukleikoen interakzioak eta proteina-lipido interakzioak....