Using electrospray ionization tandem mass spectrometry, a comparative analysis of the HaCaT keratinocyte proteins encoded by the 18th chromosome was performed before and after exposure to sodium dodecyl sulfate (25 mg/ml) and to Triton X-100 (12.5 mg/ml) in a subtoxic dose for 48 hours. Proteins were identified using the SearchGUI platform (X!Tandem and MS-GF+ search engines). In total, 1284 proteins were found in immortalized human HaCaT keratinocytes and about 75% of them were identified by two or more peptides. Were identified, that 26 proteins were encoded by genes of chromosome 18. Among these proteins, 17 were common for control cells and HaCaT cells treated with SDS. Proteins MARE2 and CTIF were identified only in control keratinocytes. Seven identified proteins encoded by genes of chromosome 18 were found only in detergent-treated keratinocytes: LMAN1, NDUV2, SPB3, VPS4B, KDSR, ROCK1 and RHG28.
Metodom tandemnoĭ mass-spektrometrii s élektrospreĭnoĭ ionizatsieĭ proveden sravnitel'nyĭ analiz belkov keratinotsitov linii NaSaT, kodiruemykh genami 18 khromosomy, do i posle vozdeĭstviia subtoksicheskikh doz dodetsilsul'fata natriia (DSN; 25 mkg/ml) i tritona X-100 (12,5 mg/ml) v techenie 48 ch. Identifikatsiiu belkov provodili s ispol'zovaniem platformy SearchGUI (poiskovye sistemy X!Tandem i MS-GF+). Vsego v immortalizovannykh keratinotsitakh cheloveka linii NaSaT obnaruzheno 1284 belka, iz nikh okolo 75% identifitsirovano po dvum i bolee peptidam. Dostoverno identifitsirovany 26 belkov, kodiruemykh genami 18 khromosomy; iz nikh 17 iavliaiutsia obshchimi dlia kontrol'nykh kletok i kletok NaSaT posle vozdeĭstviia DSN i 15 — obshchimi dlia kontrol'nykh kletok i kletok, obrabotannykh tritonom X-100. Belki MARE2 i CTIF identifitsirovany tol'ko v kontrol'nykh keratinotsitakh. Sem' identifitsirovannykh belkov, kodiruemykh genami 18 khromosomy, opredeleny tol'ko v obrabotannykh DSN keratinotsitakh, LMAN1, NDUV2, SPB3, VPS4B, KDSR, ROCK1, RHG28.
Keywords: HPLC-MS/MS; MS-GF+; SearchGUI; Triton; X! Tandem; X!Tandem; human keratinocytes of the HaCaT line; proteins of the 18th chromosome; sodium dodecylsulfat.