Isolation of an enzyme system which will catalyze the glycosylation of extensin.
Enzymes which catalyze the glycosylation of the cell wall protein extensin using uridine diphosphate l-arabinose-(14)C as a substrate are present in a crude extract prepared from suspension cultured sycamore cells (Acer pseudoplatanus L.). This enzyme system sediments when the crude extract is subjected to centrifugation at 37000g. A base hydrolysate of the product contains a mixture of hydroxyproline-arabinosides which are electrophoretically and chromatographically identical to those obtained by hydrolysis of extensin isolated from the cell wall. The hydroxyproline-rich protein used as an acceptor in the glycosylation reactions is present in the particulate fraction. In addition, evidence is presented which indicates that hydroxyproline-rich tryptic peptides prepared from the cell wall can also be used as an acceptor by this enzyme system. The presence of Mg(2+) or Mn(2+) in the reaction mixture increases the enzyme-catalyzed incorporation of arabinose into extensin by about 1.4 times. About two-thirds of the product mixture is composed of arabinose-containing compounds which have not been identified. Some of these products appear to be hydroxyproline-glycosides which have not been previously reported.
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Michigan State University-Atomic Energy Commission Plant Research Laboratory, Michigan State University, East Lansing, Michigan 48823.