scholarly journals Substrate Promiscuity: AglB, the Archaeal Oligosaccharyltransferase, Can Process a Variety of Lipid-Linked Glycans

2013 ◽  
Vol 80 (2) ◽  
pp. 486-496 ◽  
Author(s):  
Chen Cohen-Rosenzweig ◽  
Ziqiang Guan ◽  
Boaz Shaanan ◽  
Jerry Eichler

ABSTRACTAcross evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB fromHaloarcula marismortui,Halobacterium salinarum, andHaloferax mediterraneicould readily replace their counterpart fromHaloferax volcaniiwhen introduced intoHfx. volcaniicells deleted ofaglB. As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation inArchaeaand elsewhere but also for efforts aimed at transformingHfx. volcaniiinto a glycoengineering platform.

2019 ◽  
Vol 8 (27) ◽  
Author(s):  
Shiladitya DasSarma ◽  
Alexey Fomenkov ◽  
Satyajit L. DasSarma ◽  
Tamas Vincze ◽  
Priya DasSarma ◽  
...  

ABSTRACT The genomes of two extremely halophilic Archaea species, Haloarcula marismortui and Haloferax mediterranei, were sequenced using single-molecule real-time sequencing. The ∼4-Mbp genomes are GC rich with multiple large plasmids and two 4-methyl-cytosine patterns. Methyl transferases were incorporated into the Restriction Enzymes Database (REBASE), and gene annotation was incorporated into the Haloarchaeal Genomes Database (HaloWeb).


2018 ◽  
Vol 201 (2) ◽  
Author(s):  
Ronald F. Peck ◽  
Serena M. Graham ◽  
Abby M. Gregory

ABSTRACT Halophilic Archaea are a distinctive pink color due to a carotenoid pigment called bacterioruberin. To sense or utilize light, many halophilic Archaea also produce rhodopsins, complexes of opsin proteins with a retinal prosthetic group. Both bacterioruberin and retinal are synthesized from isoprenoid precursors, with lycopene as the last shared intermediate. We previously described a regulatory mechanism by which Halobacterium salinarum bacterioopsin and Haloarcula vallismortis cruxopsin inhibit bacterioruberin synthesis catalyzed by lycopene elongase. In this work, we found that opsins in all three major Halobacteria clades inhibit bacterioruberin synthesis, suggesting that this regulatory mechanism existed in the common Halobacteria ancestor. Halophilic Archaea, which are generally heterotrophic and aerobic, likely evolved from an autotrophic, anaerobic methanogenic ancestor by acquiring many genes from Bacteria via lateral gene transfer. These bacterial “imports” include genes encoding opsins and lycopene elongases. To determine if opsins from Bacteria inhibit bacterioruberin synthesis, we tested bacterial opsins and found that an opsin from Curtobacterium, in the Actinobacteria phylum, inhibits bacterioruberin synthesis catalyzed by its own lycopene elongase, as well as that catalyzed by several archaeal enzymes. We also determined that the lycopene elongase from Halococcus salifodinae, a species from a family of Halobacteria lacking opsin homologs, retained the capacity to be inhibited by opsins. Together, our results indicate that opsin-mediated inhibition of bacterioruberin biosynthesis is a widely distributed mechanism found in both Archaea and Bacteria, possibly predating the divergence of the two domains. Further analysis may provide insight into the acquisition and evolution of the genes and their host species. IMPORTANCE All organisms use a variety of mechanisms to allocate limited resources to match their needs in their current environment. Here, we explore how halophilic microbes use a novel mechanism to allow efficient production of rhodopsin, a complex of an opsin protein and a retinal prosthetic group. We previously demonstrated that Halobacterium salinarum bacterioopsin directs available resources toward retinal by inhibiting synthesis of bacterioruberin, a molecule that shares precursors with retinal. In this work, we show that this mechanism can be carried out by proteins from halophilic Archaea that are not closely related to H. salinarum and those in at least one species of Bacteria. Therefore, opsin-mediated inhibition of bacterioruberin synthesis may be a highly conserved, ancient regulatory mechanism.


2017 ◽  
Vol 199 (21) ◽  
Author(s):  
Ronald F. Peck ◽  
Alexandru M. Pleşa ◽  
Serena M. Graham ◽  
David R. Angelini ◽  
Emily L. Shaw

ABSTRACT Halophilic archaea often inhabit environments with limited oxygen, and many produce ion-pumping rhodopsin complexes that allow them to maintain electrochemical gradients when aerobic respiration is inhibited. Rhodopsins require a protein, an opsin, and an organic cofactor, retinal. We previously demonstrated that in Halobacterium salinarum, bacterioopsin (BO), when not bound by retinal, inhibits the production of bacterioruberin, a biochemical pathway that shares intermediates with retinal biosynthesis. In this work, we used heterologous expression in a related halophilic archaeon, Haloferax volcanii, to demonstrate that BO is sufficient to inhibit bacterioruberin synthesis catalyzed by the H. salinarum lycopene elongase (Lye) enzyme. This inhibition was observed both in liquid culture and in a novel colorimetric assay to quantify bacterioruberin abundance based on the colony color. Addition of retinal to convert BO to the bacteriorhodopsin complex resulted in a partial rescue of bacterioruberin production. To explore if this regulatory mechanism occurs in other organisms, we expressed a Lye homolog and an opsin from Haloarcula vallismortis in H. volcanii. H. vallismortis cruxopsin-3 expression inhibited bacterioruberin synthesis catalyzed by H. vallismortis Lye but had no effect when bacterioruberin synthesis was catalyzed by H. salinarum or H. volcanii Lye. Conversely, H. salinarum BO did not inhibit H. vallismortis Lye activity. Together, our data suggest that opsin-mediated inhibition of Lye is potentially widespread and represents an elegant regulatory mechanism that allows organisms to efficiently utilize ion-pumping rhodopsins obtained through lateral gene transfer. IMPORTANCE Many enzymes are complexes of proteins and nonprotein organic molecules called cofactors. To ensure efficient formation of functional complexes, organisms must regulate the production of proteins and cofactors. To study this regulation, we used bacteriorhodopsin from the archaeon Halobacterium salinarum. Bacteriorhodopsin consists of the bacterioopsin protein and a retinal cofactor. In this article, we further characterize a novel regulatory mechanism in which bacterioopsin promotes retinal production by inhibiting a reaction that consumes lycopene, a retinal precursor. By expressing H. salinarum genes in a different organism, Haloferax volcanii, we demonstrated that bacterioopsin alone is sufficient for this inhibition. We also found that an opsin from Haloarcula vallismortis has inhibitory activity, suggesting that this regulatory mechanism might be found in other organisms.


2016 ◽  
Vol 10 (4s) ◽  
pp. 595-600 ◽  
Author(s):  
Witold Brostow ◽  
◽  
Haley E. Hagg Lobland ◽  

The property of brittleness for polymers and polymer-based materials (PBMs) is an important factor in determining the potential uses of a material. Brittleness of polymers may also impact the ease and modes of polymer processing, thereby affecting economy of production. Brittleness of PBMs can be correlated with certain other properties and features of polymers; to name a few, connections to free volume, impact strength, and scratch recovery have been explored. A common thread among all such properties is their relationship to chemical composition and morphology. Through a survey of existing literature on polymer brittleness specifically combined with relevant reports that connect additional materials and properties to that of brittleness, it is possible to identify chemical features of PBMs that are connected with observable brittle behavior. Relations so identified between chemical composition and structure of PBMs and brittleness are described herein, advancing knowledge and improving the capacity to design new and to choose among existing polymers in order to obtain materials with particular property profiles.


2020 ◽  
pp. 34-39
Author(s):  
Aneta Antczak-Chrobot ◽  
Maciej Wojtczak

In this research paper, development of a procedure of isolation of exopolysaccharides from frost-damaged beet and an analysis of structural and chemical composition of polymers isolated from sugar beet of different origin are presented. Total acid hydrolysis degradation integrated with HPAEC-ED analysis has been utilized to confirm the monomeric composition of the separated polysaccharides. The implementation of NMR spectral analysis and SEC chromatography of the structure of exopolysaccharides has been investigated. The results demonstrate that the chemical composition and structure of exopolysaccharides depend on their origin. Typical exopolysaccharides from Central European beet roots consist mainly of glucose monomers – and they have low branched structure – about 90% of α-1,6 linkage which is typical for dextran. The exopolysaccharides isolated from Swedish beet are characterized by 50–60% fructose monomers. They contain only about 65% α-1,6 linkages. Exopolysaccharides extracted from various origin beet differ in average molecular mass. The molecular distribution is not normal.


2020 ◽  
Vol 37 (8) ◽  
pp. 7-9
Author(s):  
Israel Odede

Purpose The paper aims to critically examine the bibliographic utility as a roadmap to increase library consortia and provide an insight into a new library consortia strategy that integrates librarians into a system of sharing both resources and knowledge. Design/methodology/approach This study adopted a literature review approach with a focus on bibliographic utility as a necessary prerequisite for effective library consortia, which is a paradigm shift from the concept of individual ownership to a collective access of distributed network resources and knowledge. Findings The reviewed literature indicated that significant bibliographic utilities and integrated library systems are factors that shaped and developed consortia activities in libraries. Originality/value The bibliographic utility has limited literature, and a few published scholarly studies have combined bibliographic utility and library consortia as strategies to share resources and knowledge


2017 ◽  
Vol 21 (3) ◽  
pp. 208-223
Author(s):  
Byung Mun Lee

Purpose The purpose of this paper is to describe and analyze the rules on the formation of contracts under Korean law and the Contracts for the International Sale of Goods (CISG) in a comparative way and introduce the relevant proposed rules under the Amendment Draft of the Korean Civil Code (KCC). In addition, it attempts to compare and evaluate them in light of the discipline of comparative law. Design/methodology/approach In order to achieve the purposes of the study, it executes a comparative study of the rules as to the formation of contracts of the CISG, Korean law and the Amendment Draft of the KCC. The basic question for this comparative study is placed on whether a solution from one jurisdiction is more logical than the others and to what extent each jurisdiction has responded to protect the reasonable expectations of the parties in the rules as to the formation of contracts. Findings The comparative study finds that most of the rules under the CISG are quite plausible and logical and they are more or less well reflected in the proposals advanced by the KCC amendment committee. On the other hand, the other rules under the CISG which have brought criticisms in terms of their complexity and inconsistent case law invite us their revision or consistent interpretation. The drawbacks of the CISG have also been well responded in the Amendment Draft of the KCC. Nevertheless, it is quite unfortunate that the Amendment Draft of the KCC still has a rule that regards any purported performance with non-material alteration of the terms of an offer as an acceptance. Originality/value This study may provide legal and practical advice to both the seller and the buyer when they enter into a contract for international sales of goods. In addition, it may render us an insight into newly developed or developing rules in this area and show us how they interact with each other. Furthermore, it may be particularly useful in Korea where there is an ongoing discussion for revision of the KCC.


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