Czy białkami da się sterować?

Funkcja i aktywność białek w czasie jest bardzo ściśle regulowana w komórce. Oczywiście, człowiek nauczył się kontrolować aktywność białek (ale bez przesady). Kontrola ta odbywa się zazwyczaj przez zastosowanie odpowiednich cząsteczek, często o charakterze terapeutycznym, które pozwalają na zahamowanie/obniżenie lub podwyższenie/wywołanie białka do działania. Jednak ta forma kontroli ma pewne niedogodności – często badacze są ograniczeni przenikalnością cząsteczek przez błonę, ich czasem trwania i rozprzestrzenianiem się związków.

Pojawia się jednak iskierka nadziei dla tych, którzy chcą białka kontrolować nieco bardziej rygorystycznie. Amerykańsko- niemiecki zespół Hanny Engelke opracował metodę kontroli funkcji białek za pomocą… (werble proszę) … światła! (fanfary)

Wykorzystali oni, opublikowaną w 2010 roku przez Gautier ze współpracownikami, metodę wprowadzania modyfikowanej lizyny do sekwencji aminowkwasowej. Modyfikacja lizyny polegała na przyłączeniu do łańcucha bocznego aminokwasu cząsteczki, która będzie łatwo odchodziła pod wpływem światła (będzie inaczej mówiąc fotoczuła). Engelke ze współpracownikami otrzymała sekwencję DNA kodującą „konstrukt” składający się z fluorescencyjnego białka (eGFP), sekwencji kodującej sygnał transportu dojądrowego (NLS) i drugiego fragmentu fluorescencyjnego (mCherry). Należy wspomnieć, że sekwencja NLS zawierała mutację, która przejawiała się powstaniem kodonu STOP w miejscu, w którym powinna być lizyna (Lis29). System opracowany w 2010 roku przez Gautier i innych, zakładał, że w momencie supresji kodonu STOP, w jego miejsce wstawiana jest modyfikowana lizyna. Zatem zespół Engelke mógł albo otrzymać w wyniku translacji wyłącznie fragment eGFP (gdy nie było supresji) albo cały konstrukt z modyfikowaną lizyną w pozycji 29 (gdy zachodziła).

 

Dodatkowo, co jest bardzo ważne, wprowadzenie modyfikacji w łańcuch boczny lizyny powoduje, że sekwencja NLS zawierająca zmieniony aminokwas przestaje być wydajnym sygnałem transportu dojądrowego. Pokazują to poniższe zdjęcia z mikroskopu (na górze – przed odbezpieczeniem lizyny konstrukt jest poza jadrem, na dole – po odbezpieczeniu w jadrze).

 
MK_3

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