The hydrophobic domain of dolichyl-phosphate-mannose synthase is not essential for enzyme activity or growth in Saccharomyces cerevisiae

Dolichyl-phosphate-mannose synthase is a membrane-bound enzyme of the endoplasmic reticulum that catalyzes the formation of dolichyl phosphate mannose from dolichyl phosphate and GDP-mannose. It is an essential enzyme for growth of Saccharomyces cerevisiae, and, like other enzymes that utilize some...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1993-08, Vol.268 (22), p.16746-16753
Main Authors: Zimmerman, J.W, Robbins, P.W
Format: Article
Language:English
Subjects:
ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
ALCOHOLES
ALCOOL
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Base Sequence
BIOCHIMIE
Biological and medical sciences
BIOQUIMICA
Carbohydrate Sequence
Cloning, Molecular
CRECIMIENTO
CROISSANCE
DNA, Fungal
Enzymes and enzyme inhibitors
Escherichia coli
Fundamental and applied biological sciences. Psychology
GLICOSILTRANSFERASAS
Glycosylation
GLYCOSYLTRANSFERASE
MANNOSE
Mannosyltransferases - biosynthesis
Mannosyltransferases - genetics
MANOSA
Miscellaneous
Molecular Sequence Data
Mutagenesis
MUTANT
MUTANTES
Peptide Fragments - metabolism
Precipitin Tests
RETICULO ENDOPLASMATICO
RETICULUM ENDOPLASMIQUE
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - growth & development
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Dolichyl-phosphate-mannose synthase is a membrane-bound enzyme of the endoplasmic reticulum that catalyzes the formation of dolichyl phosphate mannose from dolichyl phosphate and GDP-mannose. It is an essential enzyme for growth of Saccharomyces cerevisiae, and, like other enzymes that utilize some form of the lipid dolichol as substrate, dolichyl-phosphate-mannose synthase contains a putative "dolichol recognition sequence" in the predicted membrane-spanning domain. To investigate the importance of this sequence in particular and the hydrophobic region in general, a series of mutants of dolichyl-phosphate-mannose synthase were constructed that contained successive deletions or mutations of the hydrophobic region, and their in vivo functions and in vitro activities were examined. While all of the mutant proteins exhibited decreased transferase activities in vitro compared to the wild-type enzyme, the sequence was not essential for growth or for protein glycosylation in S. cerevisiae. Interestingly, although deletion of the entire hydrophobic region resulted in a soluble protein, mutant proteins containing 3 or 8 hydrophobic residues at the carboxyl terminus were still membrane-associated. These mutant proteins could be released from membranes by treatment with sodium carbonate, indicating peripheral associations
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)85480-7