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Annexin VI (AnxVI) of molecular mass 68-70 kDa belongs to a multigenic family of ubiquitous Ca2+ - and phospholipid-binding proteins. In this report, we describe the GTP-binding properties of porcine liver AnxVI, determined with a fluorescent GTP analogue, 2-(or 3')-0-(2,4,6-trinitrophenyl)guanosine 5' -triphosphate (TNP-GTP). The optimal binding of TNP-GTP to AnxVI was observed in the presence of Ca2+ and asolectin liposomes, as evidenced by a 5.5-fold increase of TNP-GTP fluorescence and a concomitant blue shift (by 17 nm) of its maximal emission wavelength. Titration of AnxVI with TNP-GTP resulted in the determination of the dissociation constant (Kd) and binding stoichiometry that amounted to 1.3 uM and 1:1 TNP-GTP/AnxVI, mole/mole, respectively. In addition, the intrinsic fluorescence of the membrane- bound form of AnxVI was quenched by TNP-GTP and this was accompanied by fluo­rescence resonance energy transfer (FRET) from AnxVI Trp residues to TNP-GTP. This indicates that the GTP-binding site within the AnxVI molecule is probably located in the vicinity of a Trp-containing domain of the protein. By controlled proteolysis of human recombinant AnxVI, followed by purification of the proteolytic fragments by affinity chromatography on GTP-agarose, we isolated a 35 kDa fragment correspond­ing to the N-terminal half of AnxVI containing Trp192 . On the basis of these results, we suggest that AnxVI is a GTP-binding protein and the binding of the nucleotide may have a regulatory impact on the interaction of annexin with membranes, e.g. forma­tion of ion channels by the protein.
The aim of the study was to determine the effect of species origin of liver in the raw material composition and kind of thermal process (pasteurisation and sterilization) on the physicochemical and sensory quality of liver pate type miscellaneous sausages. Experimental material consisted of model emulsified liver pate type miscellaneous sausages produced at the P.P.H.U. “W-D” sp. z o.o. processing plant in Skwierzyna. The technological factor, which affected properties of experimental batters and the quality of final products, was the introduction in the formulation of three different species origin types of liver (porcine, poultry and rabbit liver). In this way three variants of experimental sausages liver pate type were produced. The quality of experimental sausages was assessed on the basis of the following characteristics: cooking loss, vitamin A content, sensory analyses. The following conclusions can be formulated from the presented study. The quality of liver pate is determined both by the method of preparation of liver for production (raw, blanched) and the temperature of heating (pasteurization or sterilization). It was found that the lowest cooking loss occurs in pasteurized products produced with the additions of raw poultry and rabbit liver. Preproduction blanching of liver and higher heating temperature (sterilization) have a negative effect on the analyzed quality attributes of experimental products. Out of the three experimental products the highest scores for sensory quality attributes (apart from aroma) were given to sausages produced with the addition of poultry and rabbit liver.
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