Publication detail

Magnetic Proteinase K Reactor as a New Tool for Reproducible Limited Protein Digestion

Year: 2008

Type of publication: článek v odborném periodiku

Name of source: Bioconjugate Chemistry

Publisher name: American Chemical Society

Place: Washington

Page from-to: 966-972

Titles:

Language	Name	Abstract	Keywords
cze	Magnetický reaktor s proteinázou K pro reprodukovatelné štěpení proteinů	Magnetický reaktor s proteinázou K pro reprodukovatelné štěpení proteinů	proteináza K; imobilizovaný enzym; limitované štěpení; Michaelisova konstanta
eng	Magnetic Proteinase K Reactor as a New Tool for Reproducible Limited Protein Digestion	As an aid to differentiating between the prion proteins Prpc and PrPSc, the preparation and use of immobilized Proteinase K is described. Current routine diagnosis is based on the postmortem detection of the distinct neuropathological lesion profiles of CNS and by the presence of the PK-resistant core of the prion protein isolated from brain lysates. An assay with PK immobilized to magnetic -COOH micro- and nanoparticles can offer a convenient as well as economic method. The individual immobilization steps were verified by measuring the zeta potential of the particles. The stability of the newly developed PK magnetic reactor, was highly satisfactory. The calculated values of the apparent Michaelis constant were determined from Lineweaver?Burk plots. Human growth hormone was digested using the newly prepared magnetic PK reactor and MALDI-TOF-MS analysis of the digests showed satisfactory efficiency. Controlled digestion of PrPc from the Mov mouse cell line was demonstrated with Western blot detection.	proteinase K; immobilized enzyme; limited digestion; Michaelis constant; zeta potential measurement; cellular Prion protein

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