It becomes increasingly clear that most proteins of living systems exist as components of various protein complexes rather than individual molecules. The use of various proteomic techniques significantly extended our knowledge not only about functioning of individual complexes but also formed a basis for systemic analysis of protein-protein interactions. In this study gel-filtration chromatography accompanied by mass-spectrometry was used for the interactome analysis of human liver proteins. In six fractions (with average molecular masses of 45 kDa, 60 kDa, 85 kDa, 150 kDa, 250 kDa, and 440 kDa) 797 proteins were identified. In dependence of their distribution profiles in the fractions, these proteins could be subdivided into four groups: (1) single monomeric proteins that are not involved in formation of stable protein complexes; (2) proteins existing as homodimers or heterodimers with comparable partners; (3) proteins that are partially exist as monomers and partially as components of protein complexes; (4) proteins that do not exist in the monomolecular state, but also exist within protein complexes containing three or more subunits. Application of this approach to known isatin-binding proteins resulted in identification of proteins involved in formation of the homo- and heterodimers and mixed protein complexes.
Osnovnaia massa belkov v zhivykh sistemakh funktsioniruet ne izolirovanno, a v sostave raznoobraznykh belkovykh kompleksov. Ispol'zovanie shirokogo spektra proteomnykh metodik v poslednie gody znachitel'no rasshirilo ponimanie funktsionirovaniia ne tol'ko individual'nykh belkov, no i zalozhilo osnovy sistemnogo analiza belok-belkovykh vzaimodeĭstviĭ. Nasha rabota posviashchena analizu interaktoma belkov pecheni cheloveka s primeneniem gel'-fil'tratsii, sovmeshchennoĭ s mass-spektrometrieĭ. V shesti vybrannykh fraktsiiakh lizata tkani pecheni cheloveka so srednimi molekuliarnymi massami 45 kDa, 60 kDa, 85 kDa, 150 kDa, 250 kDa, 440 kDa bylo identifitsirovano vsego 797 belkov. V zavisimosti ot profilia raspredeleniia po fraktsiiam vse identifitsirovannye belki mozhno razdelit' na chetyre gruppy: (1) odinochnye belki, nakhodiashchiesia v monomolekuliarnom sostoianii i ne uchastvuiushchie v formirovanii stabil'nykh belkovykh kompleksov; (2) belki, sushchestvuiushchie v vide gomodimerov ili geterodimerov s sorazmernymi s nimi partnerami; (3) belki, nakhodiashchiesia chastichno v monomolekuliarnom sostoianii, chastichno – v sostave belkovykh kompleksov; (4) belki, ne sushchestvuiushchie v monomolekuliarnom sostoianii, a nakhodiashchiesia tol'ko v sostave belkovykh kompleksov s chislom sub"edinits 3 i bolee. Primenenie dannogo podkhoda k ranee izvestnym izatin-sviazyvaiushchim belkam pozvolilo vydelit' takie belki, kotorye uchastvuiut kak v formirovanii gomo- i geterodimerov, tak i slozhnykh kompleksov.
Keywords: chromatographic fractionation of biological material; protein interactomics; protein mass spectrometry; protein-protein interactions; systems analysis.