Open AccessMannosyl‐phospho‐dolichol synthase from Trichoderma reesei is activated by protein kinase dependent phosphorylation in vitro
Author(s) -
Kruszewska Joanna,
Palamarczyk Grazyna,
Kubicek Christian P.
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04640.x
Subject(s) - dolichol , trichoderma reesei , atp synthase , microsome , biochemistry , phosphorylation , enzyme , protein kinase a , biology , phosphatase , extracellular , kinase , protein subunit , chemistry , biosynthesis , gene , cellulase
Summary Preincubation of microsomes from Trichoderma reesei QM 9414 with the catalytic subunit of cyclic AMP dependent protein kinase from bovine heart resulted in increased activity of the mannosylphospho‐dolichol synthase (EC 2.4.1.83, ‘Dol‐P‐Man synthase’) present in the membranes. This effect decreased upon subsequent treatment of the microsomes with alkaline phosphatase. SDS‐PAGE of microsomal proteins indicated the presence of a strong band at 32 kDa, which corresponds to the size of yeast Dol‐P‐Man synthase. Fluorography of microsomal proteins, after incubation with [γ‐ 32 P]ATP and protein kinase, revealed the incorporation of 32 P into this 32‐kDa as well as several other microsomal proteins. Dol‐P‐Man synthase activity, assayed at various times of growth, correlated with the rise and fall of the cyclic AMP pool in T. reesei . Addition of extracellular dibutyryl cyclic AMP and theophyllin, however, did not elevate the internal cyclic AMP pool in the fungus, and did not cause an effect on Dol‐P‐Man synthase activity.