Abstract
Prolyl hydroxylase, which is responsible for the hydroxylation of peptidyl proline residues, has been isolated and purified from the green alga Chlamydomonas reinhardii. The enzyme, which appears to be loosely associated with microsomal membranes, was released into solution by sonication in the presence of detergent. Purification was achieved by ion-exchange chromatography followed by affinity chromatography using the immobilized substrate poly-L-proline. Apart from its differing substrate specificity the enzyme appears to possess similar molecular characteristics to prolyl hydroxylase isolated from animal tissues: the active enzyme is a tetramer of about 240–250 kDa and nonidentical monomers of 65 and 60 kDa. The monomers are capsule shaped having a dimension of 12×7 nm.
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Abbreviations
- Da:
-
dalton
- DEAE:
-
diethylaminoethyl
- DTT:
-
dithiothreitol
- Hepes:
-
4-(2-hydroxymethyl)-1-piperazine ethanesulfonic acid
- α-KGA:
-
α-ketoglutarate
- PAGE:
-
polyacrylamide gel electrophoresis
- SDS:
-
sodium dodecyl sulfate
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Blankenstein, P., Lang, W.C. & Robinson, D.G. Isolation and characterization of prolyl hydroxylase from Chlamydomonas reinhardii . Planta 169, 238–244 (1986). https://doi.org/10.1007/BF00392320
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DOI: https://doi.org/10.1007/BF00392320