Molecular and biochemical characterisation of trehalose/maltose transportsystem of the hyperthermophilic archaeon thermococcus litoralis

The hyperthermophile Archaeon Thermococcus litoralis transports both Disaccharide Trehalose and maltose by the same transportation complex from the medium in the cell inside. This first in the group the Archaea discovered ABC transporters could be characterized in this work nearer. The besides won knowledge was compared to the information already known bacterial bandage protein of dependent ABC transporters. One of the settings of tasks of this work was in addition to the already known information common characteristics and differences of the archaeellen Trehalose/Maltose-bandage protein TMBP with that of its bacterial homolog to lay the table to the maltose / Maltodextrin-Bindeprotein MBP. That's way became on the one hand, that in the membrane anchored bandage protein cleaned. By a staining method specific for Glykoproteine turned out that the TMBP is present as a Glykoprotein. Around a three-dimensional structure of the TMBP became to determine for the X-raying's structural analysis one, after cloning and heterologer expression in E. coli, protein cleaning method establishes itself. The structure of TMBP installs, the already described bandage proteins, homologous folds sample on. In at beginning of this work to confessed malEFG-sequence section nobody could be identified to the Energetisierung of the transportation required ABC domain. Therefore, the central project was the 'search' for the ABC domain. The corresponding gene could take place kloniert and the first characteristic going out from the DNA sequence. From 1119 bp were determined for malK 372 amino acids big protein arise. The comparison of the T. litoralis of MalK protein with ABC domains from other organisms showed that all sequences typical for these proteins exist. After successful heterologer expression in E. Coli was cleaned the protein and a detailed biochemic characteristic was executed.