A comparative study of the size-heterogeneous high mannose oligosaccharides of some insect vitellins

Nordin, John H.; Gochoco, Caroline H.; Wojchowski, Don M.; Kunkel, Joseph G.

doi:10.1016/0305-0491(84)90392-4

Cited by 6 publications

(6 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Typically, the number of glucose units is reduced in insects.The principal oligosaccharide side chain on M. sexta vitellogenin lacks glucose entirely [20], whereas four species of cockroaches are known to carry small, variable quantities of the sugar [33]. The amount of glucose found in A .…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of honey bee vitellogenin

Wheeler

Kawooya

1990

Arch Insect Biochem Physiol

View full text Add to dashboard Cite

A protocol has been developed for the purification of vitellogenin from the honey bee, Apis mellifera. Purification allows for the first characterization of a vitellogenin from the large order Hymenoptera. Hymenopteran vitellogenins are unusual among insect vitellogenins in that they contain only one type of apoprotein. The honey bee vitellogenin was isolated from hemolymph of honey bee queens by a combination of density gradient ultracentrifugation, ion-exchange chromatography, and affinity chromatography. The native vitellogenin particle is a very high density glycolipoprotein containing approximately 91% protein, 7% lipid, and 2% carbohydrate. Phospholipid and diacylglycerol are the major lipid components. The equilibrium density (1.28 g/ml) is the same as that for Manduca sexta vitellogenin, which contains a much higher proportion of lipid. The covalently bound carbohydrate moiety of the particle is high in mannose. The amino acid composition of vitellogenin is similar to those of vitellogenins from other insect species. The N-terminal amino acid sequence of the apoprotein was determined, the first such sequence for any insect vitellogenin. When analyzed by sodium dodecyl sulfate (SDS)-gel electrophoresis, A. mellifera vitellogenin resolved into a single band with an apparent Mr of 180,000. Gel filtration under reducing and native conditions yielded estimated Mr values of about 300,000.

show abstract

Section: Discussionmentioning

confidence: 99%

Purification and characterization of honey bee vitellogenin

Wheeler

Kawooya

1990

Arch Insect Biochem Physiol

View full text Add to dashboard Cite

show abstract

“…Purified [2-3H]-Man Vt was converted to glycopeptides with proteinase K as described previously [7,8], and these were digested with endo-H (4 milliunits per milligram equivalent Man) in 100-500 pl (final volume) of 0.5 M MES buffer, pH 5.5 at 37°C for 72 h. The pH of the reaction mixture was then adjusted to 6.5 with NaOH and the incubation continued at 37°C for 72 h after addition of the same number of units of endo-D. The sequentid use of endo-H and endo-D ensures that all of the oligosaccharides present are liberated regardless of the number of mannosyl residues they contain [19,20].…”

Section: Preparation Of [3h]-man-labeled Oligosaccharidesmentioning

confidence: 99%

“…The latter two are more highly glycosylated on both a mass and molar basis than the M, 50,000 subunit. Because B. germunicu Vt is 5% carbohydrate and the major oligosaccharide species is MangGlcNAc2 [7,8], the average number of chains on the subunits could be calculated. Each M, 50,000 subunit contains an average of a single oligosaccharide chain; the M, 95,000, four chains; and the M, 102,000, three chains.…”

Section: Constitution Of Vt Subunitsmentioning

confidence: 99%

“…Knowing what regulates the timing of the limited proteolysis at day 4 and which structural features of the Vt molecule contribute to the selectivity of this event is important for understanding how Vt utilization is regulated. Although insect Vt's contain covalently bound phosphate [2,6,7 and high Man-type oligosaccharides [8,9], their fates during embryogenesis, as well as any functions they might have in controlling Vt degradation have not been investigated. Although, this day-4 event may itself induce structural modifications that mark Vt for utilization, removal of saccharide or phosphate residues could facilitate proteolysis of selected domains of the molecule by inducing conformational changes or deshielding critical amino acid residues.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Yolk hydrolase activities associated with polypeptide and oligosaccharide processing of Blattella germanica vitellin

Purcell

Kunkel

Nordin

1988

Arch Insect Biochem Physiol

Self Cite

View full text Add to dashboard Cite

Various aspects of the processing of Blattella germanica vitellin (Vt) in the oocyte and egg have been investigated. Employing subunit specific antibodies, the precursor product relationships among t h e subunits of this Vt have been determined. After endocytosis of Vt by the oocyte, t h e M, 160,000 subunit Vt is cleaved to products of M, 95,000 and M, 50,000. In association with an unprocessed M, 102,000 peptide, these form the subunits of the Vt of freshly ovulated eggs. Between 4 and 5 days post ovulation (at 3OoC), all three subunits of Vt are again processed proteolytically before use by the embryo. Although Vt's high mannose-type oligosaccharides are trimmed during embryogenesis, their modification occurs subsequent to the day 4-5 proteolysis, precluding the possibility that changes in oligosaccharide content or structure contribute to regulating this second proteolytic event.Although the predominant oligosaccharide of Vt is Man9GlcNAc2, the M, 50,000 subunit of egg-borne Vt contains a much higher proportion of Man6GlcNAc2 than t h e other two subunits; therefore, this portion of the precursor vitellogenin must be more accessible to the processing mannosidases of the endoplasmic reticulum during its biosynthesis. A microtechnique for aspirating the yolk from individual eggs in an ootheca permits its isolation free of contamination by embryonic tissue. With this procedure, the specific activity profiles of exo-a-rnannosidase, exo-0-Nacetylgl ucosam i n idase, a-glucosidase and acid phosphatase were monitored during the first 6 days after ovulation, and some of their properties were also determined. Expression of the acid phosphatase and exo-P-N-acetyl- glucosarninidase activities coincide with the day 4-5 proteolysis, while arnannosidase remains relatively constant throughout the first 6 days.Functions for these enzymes and the oligosaccharides of Vt during Vt storage and utilization are proposed.

show abstract

“…A major product of very similar size results from labeling in intact fat body when glycosylation is inhibited by tunicamycin [13]. It therefore seems likely that the two genes code for identically sized mRNAs and primary translation products that undergo size differentiation by addition of the carbohydrate moiety [14].…”

Section: The Locust Vitellogenin Genesmentioning

confidence: 98%

Juvenile hormone-regulated locust vitellogenin genes: Lack of expression after transfer intoDrosophila

Wyatt

Kanost

Locke

et al. 1986

Arch. Insect Biochem. Physiol.

View full text Add to dashboard Cite

In Locusta rnigratoria, vitellogenin (Vg) is normally produced only in adult female fat body under stimulation by juvenile hormone (JH). This permits study of a) programming of genes for expression and b) modulation of expression by JH. From L. rnigratoria genomic libraries, two Vg genes (A and B) have been cloned. Coding regions encompass 10-12 kbases of DNA, contain introns, and hybridize with 6,300 nucleotide mRNAs. Although the 5'-exons, coding for signal peptides, are similar in sequence, the remaining coding sequences have distinct restriction maps and show no crosshybridization. Upstream DNA from the two genes has some sequence similarity, corresponding to potential regulatory regions. Dot hybridization assays of mRNAs A and B in locust fat body after induction by methoprene show coordinate expression of the two Vg genes and also a third, unidentified JH-regulated gene. In the hope of providing a system for identification of control sequences, P element-mediated transformation has been used to transfer locust DNA into Drosophila. From Vg gene B, a central block was deleted, to give a mini-Vg gene comprising 5' and 3' terminal coding sequences plus upstream flanking DNA. This was incorporated into the P element vector Carnegie 20 and injected into Drosophila embryos. Three transformed fly lines were obtained in which the locust mini-Vg gene was integrated at different chromosomal sites. On Northern blots of fly RNA, however, no expression of the locust sequences could be detected, even though the accompanying rosy gene was expressed. This suggests that the locust regulatory sequences were not recognized in Drosophila, and current effort i s directed toward developing a homologous locust transformation system for expression assay of cloned JH-regulated genes.

show abstract

A comparative study of the size-heterogeneous high mannose oligosaccharides of some insect vitellins

Cited by 6 publications

References 35 publications

Purification and characterization of honey bee vitellogenin

Purification and characterization of honey bee vitellogenin

Yolk hydrolase activities associated with polypeptide and oligosaccharide processing of Blattella germanica vitellin

Juvenile hormone-regulated locust vitellogenin genes: Lack of expression after transfer intoDrosophila

Contact Info

Product

Resources

About