Isolation and1H-NMR spectroscopic identification of the glucose-containing lipid-linked precursor oligosaccharide ofN-linked carbohydrate chains

The lipid-linked precursor ofN-type glycoprotein oligosaccharides was isolated from porcine thyroid microsomes after in cubation with UDP[³H] Glucose. The carbohydrate was released from dolichol pyrophosphate by mild acid hydrolysis, purified by gel filtration and characterized by 500-MHz¹H-NMR spectroscopy in combination with enzymatic degradation. The parent oligosaccharide was found to be Glc₃Man₉Glc-NAc₂. The three glucose residues are present in the linear sequence Glcα1-2Glα1-3 Glc, the latter being α(1-3)-linked to one of the mannose residues. In order to establish the branch location of the triglucosyl unit, the parent compound was digested with jack-bean α-mannosidase. The oligosaccharide product was purified by gel filtration, and identified by¹H-NMR as Glc₃Man₅GlcNAc₂ lacking the mannose residues A, D₂, B and D₃. Therefore, the structure of the precursor oligosaccharide is as follows: $$\begin{gathered} c b a D_1 C 4 \hfill \\ Glc\alpha 1 - 2Glc\alpha 1 - 3Glc\alpha 1 - 3Man\alpha 1 - 2Man\alpha 1 - 2Man\alpha 1 \hfill \\ 3 \swarrow 3 2 1 \hfill \\ Man\alpha 1 - 2Man\alpha 1 Man\beta 1 - 4GlcNAc\beta 1 - 4GlcNAc \hfill \\ D_{2 } A 3 6 \hfill \\ Man\alpha 1 \hfill \\ 6 \hfill \\ Man\alpha 1 - 2Man\alpha 1 \nwarrow 4 \hfill \\ D_3 B \hfill \\ \end{gathered} $$      

An epitope on C4 β light (L) chains detected by human anti-Rg; its relationship with β chain polymorphism and MHC associations

Two out of ten Rg-specific antisera tested contain a third antibody specific for the β chain of C4. Analysis of the β chains of 66 unrelated individuals by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the epitope detected is located exclusively on the light (L) β chain. A strong, but incomplete, association between the β chain epitope and the expression of the Rg: 2 determinant on the α chain of the same protein was also observed. While H (heavy) and L β chains were not associated with a particular C4 isotype, previously unrecorded associations of β chain polymorphism with theDR locus have been established.     

Imperfect oviposition decisions by the pitcher plant mosquito (Wyeomyia smithii)

Summary Wyeomyia smithii mosquitoes distribute their eggs across available oviposition sites (water-holding pitcher plant leaves) of varying quality. I experimentally examined responses to three components of site quality: conspecific larval density, larval density of the pitcher plant midge,Metriocnemus knabi, and pitcher size. Responses to larval treatments were complex and apparently suboptimal. Although mosquito larval performance is better in leaves with fewer conspecific and more midge larvae, females did not lay more eggs in such pitchers. Instead, more eggs were laid in experimental pitchers containing either midge or mosquito larvae, but fewer eggs in pitchers with neither or both. More eggs were laid in larger pitchers, which tend to accumulate more resources and dry out less often. Therefore, although the oviposition decisions made were suboptimal, they were better than random.     

Oviposition and feeding preferences of a flower-feeding weevil,Coelocephalapion aculeatum, in relation to conspecific damage to its host-plant

The oviposition and feeding preferences ofCoelocephalapion aculeatum Fall (Coleoptera: Apionidae), a host specific florivore ofMimosa pigra L. (Mimosaceae), were studied in relation to conspecific damage to its hostplant. Adults ofC. aculeatum cease ovipositing in inflorescences when the egg load reaches a number consistent with the larval carrying capacity of the inflorescence. The basis for this oviposition deterrence was examined by offering inflorescences damaged by adult feeding alone, larval feeding alone and a combination of adult feeding and oviposition. Adults preferred to oviposit on inflorescences which are not damaged by either adult feeding, larval feeding, or oviposition. No evidence for the existence of an oviposition deterring pheromone (ODP) was found. I suggest that the ability of a single host inflorescence to support the development of many larvae causes selection for the use of these oviposition deterring cues which can convey more quantitative information about the level of previous infestation than can ODPs. Adults fed a similar amount on damaged compared to undamaged inflorescences. These results assisted in the design of host range testing trials and allows predictions to be made about the effectiveness of this insect as a biological control agent.     

Biology and Host Range of the Green-seed Weevil, Sibinia fastigiata , for Biological Control of Mimosa pigra

Aspects of the biology and host range of Sibinia fastigiata Clark (Coleoptera: Curculionidae) were studied to assess its safety for release in Australia as a biological control agent of the weed Mimosa pigra L . (Mimosaceae) . Larvae feed on the seeds and adults on open flowers of their host . Adults oviposit on to immature seeds 3 mm long or less and hence seeds of this length and maturity were used in the host range tests and for rearing . Females are shown to avoid previously attacked seeds enhancing their effectiveness as seed destroyers . Survival of adults was higher when provided with open flowers . The host range was determined using laboratory control - choice oviposition tests on excised plant material and , in the field in the native range , no - choice oviposition tests on living plants , surveys of adults on plants , and breeding of insects from pods of plants of various legume species . The control - choice oviposition tests employed a new design in which the control plant alone was offered to the insects followed by a choice of test plants species . Other than M. pigra, only one plant species was acceptable for oviposition , the closely related M. asperata. Larval development also occurs on M. asperata and this host is occasionally used in the field . This insect was approved for release in March 1997 .     

Receptor choice determinants in the envelope glycoproteins of amphotropic, xenotropic, and polytropic murine leukemia viruses.

The envelope glycoproteins (SU) of mammalian type C retroviruses possess an amino-terminal domain of about 200 residues, which is involved in binding a cell surface receptor. In this domain, highly conserved amino acid sequences are interrupted by two segments of variable length and sequence, VRA and VRB. We have studied the role of these variable regions in receptor recognition and binding by constructing chimeric molecules in which portions of the amino-terminal domains from amphotropic (4070A), xenotropic (NZB), and polytropic (MCF 247) murine leukemia virus SU proteins were permuted. These chimeras, which exchanged either one or two variable regions, were expressed at the surface of replication-defective viral particles by a pseudotyping assay. Wild-type or recombinant env genes were transfected into a cell line producing Moloney murine leukemia virus particles devoid of envelope glycoproteins in which a retrovirus vector genome carrying an Escherichia coli lacZ gene was packaged. The host range and sensitivity to interference of pseudotyped virions were assayed, and we observed which permutations resulted in receptor switch or loss of function. Our results indicate that the determinants of receptor choice are found within the just 120 amino acids of SU proteins. Downstream sequences contribute to the stabilization of the receptor-specific structure.