Elaboration and introduction of the method for determination of some proteins

Abstract

Core protein of aggrecan has a significant share on the correct function of articular cartilage. Its lack or structural failure could be the reason for the disfunction of the cartilage. The culture of chondrocytes taken from a pork articular cartilage was used for the study of aggrecan production. The monolayer culture method offers the model system which has enabled us to watch the aggrecan production into growth medium. The aggrecan synthesis was stimulated in the media with addition of L-methionin, L-serin and sodium selenite pentahydrate. Methionin and serin are antecedents of sulphur amino acid of cysteine, whose role is incredibly important for the correct function of core protein. Growth media and chondrocytes were analysed with the help of the automatic amino acids analyzer unit after acid or oxidative hydrolysis. The analyse established the amino acid representation. The main attention was paid to cysteine. The changing concentrations of this amino acid were showing if the antecedents in the addition are used for its production and, therefore, if it is possible to stimulate the production of core protein with these antecedents. The results are discussed in the conclusion of this thesis. The next step should be the detection of the concentration of synthesized aggrecan by the immunological method. Presently this method is very expensive. Therefore, the method of setting the core protein of aggrecan with the help of suitable amino acid was used for the first tests