|
KMID : 0374919920130040461
|
|
Inje Medical Journal
1992 Volume.13 No. 4 p.461 ~ p.478
|
|
|
Purification and Biological properties of Adenosine Deaminase Complexing Protein From Bovine Kidney
|
|
|
|
|
Abstract
|
|
|
ADCP (adenosine deaminase complexing protein) was prepared and purified from bovine kidney. Its purification was done by the treatment of Triton X-100, followed by precipitation with sodium sulfate. ADA-Sepharose affinity chromatography and
finally
Sephadex G-100 gel filteration. The purified ADCP thus obtained is supposed to exist as a homodimer of molecular weight about 200 KD on SDS-polyacrylamide gel electrophoresis. The presence of ADCP caused a slight decrease in Km and Vmax value for
adenosine, but the ADA (adenosine deaminase) binding with ADCP did not modify its behavior of optimal pH and temperature. No effects of varying metal ions, purine metabolites and its derivatives were observed on the activities of ADA in the
presence of
ADCP, except 6-chloropurine riboside inducing 27% decrease in activity and Fe2+ enhancing 29% of its activity. The binding of ADA with ADCP was optimal at 1.7:1 molar ratio of ADA-ADCP with no effect of the addition of varying metal ions or
purine
metabolites except the guanosine and 2.6-diaminopurine, both causing aggragating complex with their concentration.
The effect of ADCP on catalytic activity of ADA was studies. The slight elevation (up to 13%) of ADA catalytic activity was observed at the adenosine contents of 0.1-0.2mM and ADCP/ADA molar ratio of 1:1~1:10, while the decreased in its activity
was
shown at the same range of adenosine contents, but ADCP/ADA molar ratio of 2:1~10:1. The lower activities of ADA were also observed at lesser contents(0.01~0.05mM) of adenosine and the same molar ratio of ADCP/ADA. Thus it was shown that ADCP
interacted
with ADA. And the ADA activity tends to decrease at the physiological concentration of adenosine (about 0.01mM). These results show that ADCP might play a role in the regulation of ADA activity.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|