Enzymatic isolation of cells from bone: cytotoxic enzymes of bacterial collagenase
The enzymatic isolation of cells from fetal rat calvaria is most effectively achieved with crude Clostridium histolyticum collagenase. However, this bacterial collagenase damages the cells during the digestion of the tissue. We have used cell density, as measured by isopycnic centrifugation on polysucrose gradients, as an indicator of cell damage. There are at least two enzymes in crude bacterial collagenase capable of damaging the cells in this tissue. One of these is clostripain that has been well characterized.The other cytotoxic enzyme is uncharacterized, and its effects are not evident until the clostripain activity has been inhibited by ..cap alpha..-tosyl-lysyl chloromethane. The apparent activity of this second enzyme can be inhibited by withholding magnesium from the digestion medium and by increasing the potassium concentration of the digestion medium.
- Research Organization:
- Univ. of Rochester, NY
- DOE Contract Number:
- AC02-76EV03490
- OSTI ID:
- 5212294
- Journal Information:
- Am. J. Physiol.; (United States), Vol. 240
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PEPTIDE HYDROLASES
BIOCHEMICAL REACTION KINETICS
ENZYME INHIBITORS
BIOCHEMISTRY
BONE CELLS
CATIONS
CENTRIFUGATION
CHEMICAL COMPOSITION
CLOSTRIDIUM
COLLAGEN
ENZYME ACTIVITY
FETUSES
MAGNESIUM
POTASSIUM
RATS
SKELETON
TOXICITY
ALKALI METALS
ALKALINE EARTH METALS
ANIMAL CELLS
ANIMALS
BACTERIA
BODY
CHARGED PARTICLES
CHEMISTRY
CONNECTIVE TISSUE CELLS
ELEMENTS
ENZYMES
HYDROLASES
IONS
KINETICS
MAMMALS
METALS
MICROORGANISMS
ORGANIC COMPOUNDS
ORGANS
PROTEINS
REACTION KINETICS
RODENTS
SCLEROPROTEINS
SEPARATION PROCESSES
SOMATIC CELLS
VERTEBRATES
550200* - Biochemistry