Glycosyl transfer to an acceptor lipid from insects

Allué, Luis Alberto Quesada; Belocopitow, Enrique; Maréchal, Luis R.

doi:10.1016/0006-291x(75)90486-6

Cited by 21 publications

(12 citation statements)

References 13 publications

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“…This extract was about 1/3rd as active in forming Dol-P-Mann (cpm//Jg protein) as the corresponding microsome-enriched preparations [ 18]. Since no bactoprenyl derivative was detected by thin-layer chromatography [15] it was concluded that bacterial contamination from the digestive tract was negligible.…”

Section: Enzyme Preparationmentioning

confidence: 87%

“…Dol-P was detected by the enzymatic test in [15]. (A) Non-hydrolyzed; (B) treated with 0.01 M HC1 for 30 min at 100°C; (C) strips 7,8, 9 and 10 (corresponding to Dol-P) of experiment B were washed with toluene (to remove the scintillator), the silica gel was scraped off and extracted with 0.6 N HC1 in chloroform/methanol (2/1) [15 ]. The extract was then chromatographed on paper using solvent E and radioactivity was located as above.…”

Section: Atpmentioning

confidence: 99%

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Phosphorylation of dolichol by insect enzymes

Allué¹

1979

FEBS Letters

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Section: Enzyme Preparationmentioning

confidence: 87%

Section: Atpmentioning

confidence: 99%

Phosphorylation of dolichol by insect enzymes

Allué¹

1979

FEBS Letters

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“…[37]. Liver Dol-P-Glc (labeled with 14C or 3H in the glucose residue) and insect dolichyl-P were prepared as described [32].…”

Section: Methodsmentioning

confidence: 99%

“…It has been recently reported that dolichol [30,31], dolichyl phosphate [32] and dolichyl derivatives [33,34] occur in pluricellular invertebrates, but little is known about the mechanisms involved in protein glycosylation in those metazoan phyla [35].…”

Section: ~_ _ _mentioning

confidence: 99%

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Lipid‐Bound Oligosaccharides in Insects

Allué

Belocopitow

1978

European Journal of Biochemistry

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Membrane preparations from immature stages of the fruit fly Ceratitis capitata catalyze the transfer of mannose from GDP-['4C]mannose into lipid-linked oligosaccharides. These compounds behave as polyprenyl derivatives and their formation is stimulated by the addition of an acidic glycolipid fraction isolated from insects.The mannose-labeled oligosaccharides are attached to the poly-isoprenol by a pyrophosphoryl linkage and can be released by mild acid hydrolysis. The trisaccharide lipid has been partially characterized. The results indicate that the compound is polyprenyl-pyrophosphate-N,N'-diacetylchitobiose-mannose.Incubation of dolichyl phosphate [14C]mannose or lower 14C-labeled oligosaccharide lipids with unlabeled GDP-mannose and the insect enzyme leads to the labeling of a higher lipid-bound oligosaccharide.When UDP-N-a~etyl['~C]glucosamine was incubated with insect membranes a '4C-labeled chitobiosyl lipid was synthesized. If unlabeled GDP-mannose was also present, the I4C label appeared in the trisaccharide and higher oligosaccharide lipids.Preliminary evidence indicates that the insect polyprenyl oligosaccharides described here might participate in glycoprotein biosynthesis.The role of polyprenylphosphates as glycosyl carriers in eucaryotic tissues is now well established [l-61. Evidence is being accumulated showing that dolichyl-bound saccharides are intermediates in the glycosylation of proteins in vertebrate [7 -141 plant [15,16] and fungal [17-211 systems. First, GlcNAc from UDP-GlcNAc is transferred to dolichyl-P. Then, N-acetylglucosamine and mannose are sequentially added from their respective sugar nucleotides yielding Dol-P-P-(GlcNAc)2-Man [22] which can undergo additional mannosylations [S, 11,221. Some of these mannose residues probably came from Dol-PMan which, in turn, is formed through the reaction of dolichyl-P with GDP-Man 123 -251. The last step is the transfer of the oligosaccharide from dolichyl-P-P derivatives to the polypeptide chains. The transfer of ['4C]mannose-labeled oligosaccharides has been described in yeast [26] and in vertebrate systems [8 -l l , 27,281.The amino acid involved in the protein-saccharide linkage is presumably asparagine because the sequence N,N'-diacetylchitobiose-mannose has always been found linked to it [29]. ~_ _ _Abbreviations. GlcNAc, N-acetylglucosamine; Dol, dolichyl ; Man, mannose; dolichyl-P. dolichyl moiiophosphate; dolichyl-P-P, dolichyl diphosphate.Enzyme. Streptomyes griseus protease or streptococcal proteinase (EC 3.4.22.10).It has been recently reported that dolichol [30,31], dolichyl phosphate [32] and dolichyl derivatives [33,34] occur in pluricellular invertebrates, but little is known about the mechanisms involved in protein glycosylation in those metazoan phyla [35].We previously reported the transfer of glucose, mannose and N-acetylglucosamine from their respective nucleotides to an endogenous polyprenyl phosphate catalyzed ip2 vitro by an insect extract [33,36]. These reactions could be stimulated by the addition of insect or pig li...

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Characterization of mannosyl transferases during the pupal instar of Stomoxys calcitrans (L)

Mayer

Chen

DeLoach

1983

Arch Insect Biochem Physiol

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Particulate fractions (10,OOOg) from pupae of Stomoxys calcitrans transfer[14C]-mannose from CDP-[14C]-mannose to dolichol monophosphate and proteins. production of the mannosyl lipid was inhibited by Mn2+, UDP, GMP, GDP, and EDTA. The insect growth regulator diflubenzuron had no effect on mannosyl transferase activity. Dolichol monophosphate and Mg2+ stimulated mannosyl transferase activity. The mannosyl lipid product was identified as mannosyl-phosphoryl-dolichol (Man-P-Dol)*. The apparent K , and V, , , values for the formation of Man-P-Do1 using GDP-[14C]-Man while holding dolichol phosphate constant were 2.4 k 0.9 p M and 9.4 k 2.3 pmol Man-P-Do1.min-l.mg-l protein, respectively. The apparent K , and V , , , values using dolichol phosphate while holding GDP-Man constant were 2.2 f 1.2 p M and 18.5 f 1.7 pmol Man-P-Dol.min-'.mg-' protein.

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Glycosyl transfer to an acceptor lipid from insects

Cited by 21 publications

References 13 publications

Phosphorylation of dolichol by insect enzymes

Phosphorylation of dolichol by insect enzymes

Lipid‐Bound Oligosaccharides in Insects

Characterization of mannosyl transferases during the pupal instar of Stomoxys calcitrans (L)

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