Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: EC:1.14.11.2 (
prolyl hydroxylase
)
1,814
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
In the purification of
proline hydroxylase
by affinity chromatography on poly(L-proline)-Sepharose it was found earlier that two other components,
profilin
and the complex
profilin
-actin, also bind with high affinity to this matrix. We have exploited this observation to develop a rapid procedure for the isolation of
profilin
and
profilin
-actin complexes in high yields directly from high-speed supernatants of crude tissue-extracts. Through an extensive search for elution conditions, avoiding poly(L-proline) as the desorbant, we have found that active proteins can be recovered from the affinity column with a buffer containing 30% dimethyl sulphoxide. Subsequent chromatography on hydroxylapatite separates free
profilin
and the two isoforms of profilactin,
profilin
-actin beta and
profilin
-actin gamma. The
profilin
-actin complexes produced this way have high specific activities in the DNAase-inhibition assay, give rise to filaments on addition of Mg2+, and can be crystallized. From the isolated
profilin
-actin complexes the beta- and gamma-actin isoforms of non-muscle cells can easily be prepared in a polymerization competent form. Pure
profilin
is either obtained from an excess pool present in some extracts or by dissociation of
profilin
-actin complexes and removal of the actin.
...
PMID:The use of poly(L-proline)-Sepharose in the isolation of profilin and profilactin complexes. 319 57
