Collagen enzyme studies

Studies of the enzyme content of cells frequently involve the use of coarse tissue samples of either animal or plant origin. In such cases some preliminary dissection of the tissue may be necessary to isolate the relevant tissue components and remove unwanted structural material such as collagen, cellulose, etc., before moving on to the more critical disruption of the cells. Sometimes it is possible to use the technique of tissue culture to provide pure cell preparations for subsequent studies. 

In in vitro studies penicillamine inhibited angiotensin-con-verting enzyme (ACE) and carboxypeptidase (930). Penicillamine interferes with the functions of the copper-containing enzyme ceruloplasmin, and some of the penicillamine- and copper-containing complexes formed in vivo have a superoxide dismutase effect (931). In patients with scleroderma, penicillamine normalized collagen metabolism, by inhibiting beta-galactosidase activity (932). 

Equilibrium Sorption Procedure. The sorption (binding) of 8-galactosidase by the various modified and unreacted collagen membranes was measured by the method of equilibrium sorption (18, 13). In this procedure, collagen membrane was impregnated with the purified enzyme (8-galactosidase, J2. coli K12) as a function of enzyme bath concentrations. Enzyme solutions containing 0.023 0.012, 0.0074, 0.0047) 0.0031 and 0.0021 pmole/ml of enzyme in phosphate buffer (0.02H) at pH 7-0 were employed as the sorption baths. All sorption studies were carried out at 4°C for a twenty four hour period. Transient state sorption data established that the system had equilibrated within the twenty four hour period. 

---