Proteomic analysis of the nuclear fraction is of great importance, since many cellular processes are initiated in the nucleus. Refinement and choice of experimental procedures for cell lysate fractionation and parameters for mass spectrometric detection and data processing continue to be of current interest. The mass spectrometry analysis presented here was tested on human cell lines derived from different tissues: HL-60 (peripheral blood); HepG2 (liver); EA.hy926 (vascular endothelium). High reproducibility of results and their consistency with biological properties of the objects under study were demonstrated. The use of cells of different types made it possible to reveal a set of 16 proteins whose LFQ-values allow for the discrimination between proteome fractions regardless of cell origin. Also, a set of 16 proteins is suggested which are associated with individual characteristics of cell lines regardless of cell fraction. These protein panels can serve as parameters to verify the proteomic analysis done was of sufficient quality, in particular as indicators of successful fractionation of cell or tissue lysate.
Issledovanie proteoma iadernoĭ fraktsii iavliaetsia vazhnoĭ zadacheĭ, poskol'ku bol'shaia chast' protsessov, proiskhodiashchikh v kletke, initsiiruetsia v iadre. Razrabotka i vybor usloviĭ fraktsionirovaniia kletochnykh lizatov i parametrov mass-spektrometricheskogo detektirovaniia i obrabotki dannykh v nastoiashchee vremia prodolzhaiut ostavat'sia aktual'nymi. Predstavlennyĭ v rabote mass-spektrometricheskiĭ analiz aprobirovan na liniiakh kletok, proiskhodiashchikh iz raznykh tkaneĭ: HL-60 (perifericheskaia krov'), HepG2 (pechen'), EA.hy926 (éndoteliĭ sosudov). Prodemonstrirovana vysokaia vosproizvodimost' rezul'tatov i ikh soglasovannost' s biologicheskimi svoĭstvami issledovannykh ob"ektov. Blagodaria ispol'zovaniiu raznogo tipa kletochnykh liniĭ vyiavlen nabor iz 16 belkov, ch'i kolichestvennye pokazateli (LFQ – Label-free Quantification Intensity) pozvoliaiut diskriminirovat' iadernuiu i tsitozol'nuiu fraktsii vne zavisimosti ot proiskhozhdeniia kletok. Takzhe predlozhen nabor iz 16 belkov, ch'i LFQ sviazany s individual'nymi priznakami kletochnykh liniĭ vne zavisimosti ot fraktsii. Éti paneli belkov mogut ispol'zovat'sia kak indikatory uspeshnogo fraktsionirovaniia kletochnogo ili tkanevogo lizata.
Keywords: EA.hy926; HL-60; HepG2; nuclear protein fraction; proteomic profiling.