Presentations & Public Speaking

Biomolecular engineering of human lectin galectin-2

Biomolecular engineering of human lectin galectin-2
of 1
All materials on our website are shared by users. If you have any questions about copyright issues, please report us to resolve them. We are always happy to assist you.
Related Documents
  understandingof biophysical propertiesof glycoproteinsandprotein – glycan conjugates for biotechnological purpose. References 1.  Shental-Bechor,D.andLevy,Y.: Effectofglycosylationonproteinfolding:acloselookat thermodynamic stabilization, Proc. Natl. Acad. Sci. USA, 105,  8256-8261 (2008).2.  Choi, Y., Lee, J.H., Hwang, S., Kim, J.K., Jeong, K., and Jung, S.:  Retardation of theunfolding process by single N-glycosylation of ribonuclease A based on moleculardynamics simulations. Biopolymers.  89.,  114-123 (2008).3.  Wyss, D.F. and Wagner, G.:,  The structural role of sugars in glycoproteins. Curr. Opin.Biotechnol.,  7.,  409-416 (1996).doi:10.1016/j.jbiosc.2009.08.178 BE-O11Discovery of extracellular respiration of NADH and its applicationin novel NADH regeneration Chong Zhang, 1 Yo Hirose, 1,2 Xi Wu, 1 Kun Ma, 1 Ichiro Okura, 2 andXin-Hui Xing 1 Department of Chemical Engineering, Tsinghua University, Bejing, China 1 Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Tokyo, Japan 2 Soluble substrates can be utilized extracellularly by extracellularrespiration. But no study has showed NADH could be respired outsidethe cells. The present work have interestingly discovered that NADHcould be extracellularly respired by many organisms.TheextracellularNADHoxidase activities(ENOAs)ofwholecellsof  Enterobacter aerogenes ,  Pseudomonas putida  and  Bacillus cereus  were0.1,0.5and1.2U/OD 600 ,respectively.Whenthewholecellsoftheabovethree strains were treated with proteinase K, the ENOAs were lost,suggesting that the NADH oxidase was located outside of the cells. ToclarifytheextracellularNADHoxidizationsystem,thegeneralsecretionpathwayIIwiththefunctionforproteinsecretionwasknocked-outforanalyzing its influences on the ENOAs. Moreover, the extracellularNADH oxidase was purified and sequenced. And the function of thetarget enzyme was confirmed through its overexpression.As far as we know, this is the first report on the discovery of extracellularrespirationofNADH.TheextracellularNADHoxidationisconsidered as a promising tool for developing novel NADH regenera-tion system.Acknowledgments: This work was supported by the Project of theNatural Science Foundation of China (Grant No. 20806046) and theNational Basic Research Program of China (Grant No. 2009CB724702). doi:10.1016/j.jbiosc.2009.08.179 BE-O12Biomolecular engineering of human lectin galectin-2 Hui Wang, 1 Lizhong He, 1 Martin Lensch, 2 Hans-Joachim Gabius, 2 Conan J. Fee, 3 and Anton P.J. Middelberg 1 The University of Queensland, Australian Institute for Bioengineering andNanotechnology, Centre for Biomolecular Engineering, St Lucia,QLD 4072, Australia 1 Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University, 80539 Munich,Germany 2 and Department of Chemical and Process Engineering,University of Canterbury, Christchurch 8140, New Zealand 3 The activity of galectins as potent regulators of cell growth andadhesion, e.g. in malignancy and autoimmune diseases, make themattractive drug candidates (1). In this work, we demonstrate thatthese endogenous lectins can be transformed into pharmaceuticallystable forms, using human galectin-2 (Gal2) as a proof-of-conceptexample. We constructed three mutants of Gal2 (C57A, C57M andC57S) by introducing mutations at the site of one of the two Cysresidues. Only the C57M variant was expressed in  E. coli  in highlysoluble form. Mutant C57M retained its binding ability to lactose,facilitating a single-step affinity purification using lactose-agarose.The modified protein showed no detectable aggregation followingthree weeks of storage, in contrast to significant aggregation for thewild type protein. The C57M mutation enabled site-specificchemical modification as exploited by conjugation with poly-ethylene glycol at the remaining sulfhydryl group (Cys75). Ionexchange chromatography was used to separate homogenous PEG-Gal2 from the reaction solution.The results thus demonstrate the feasibility of combined geneticand chemical modifications to enhance the suitability of a humanlectinasapharmaceuticallyrelevantprotein,inadditiontotheknownphysiological benefits of PEGylation. Reference 1.  Gabius, H.-J.:  The sugar code. Fundamentals of glycosylation. Wiley-VCH. Weinheim(2009).doi:10.1016/j.jbiosc.2009.08.180 BE-O13Construction of a novel detection system for protein – proteininteractions using yeast G-protein signaling  Nobuo Fukuda, 1  Jun Ishii, 2 Tsutomu Tanaka, 2 and Akihiko Kondo 1 Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan 1 and Organization of  Advanced Science and Technology, Kobe University, Kobe, Japan 2 In the current study, we report the construction of a novelsystem for the detection of protein-protein interactions using yeastG-protein signaling. It is well established that the G-protein gammasubunit (Ggamma) is anchored to the inner leaflet of the plasmamembrane via lipid modification in the C-terminus, and that thislocalization of Ggamma is required for signal transduction. In oursystem, mutated Ggamma (Ggamma cyto ) lacking membrane locali-zation ability was genetically prepared by deletion of the lipidmodification site. Complete disappearance of G-protein signal wasobserved when Ggamma cyto  was expressed in the cytoplasm of yeast cells from which the endogenous Ggamma gene had beendeleted. In order to demonstrate the potential use of our system, weutilized the  Staphylococcus aureus  ZZ domain and the Fc portion of human immunoglobulin G (IgG) as a model interaction pair. Todesign our detection system for protein-protein interaction, the ZZdomain was altered so that it associates with the inner leaflet of theplasma membrane, and the Fc part was then fused to Ggamma cyto .The Fc-Ggamma cyto  fusion protein migrated towards the membranevia the ZZ-Fc interaction, and signal transduction was thereforeS61  Abstracts / Journal of Bioscience and Bioengineering 108 (2009) S57  – S74
Similar documents
View more...
Related Search
We Need Your Support
Thank you for visiting our website and your interest in our free products and services. We are nonprofit website to share and download documents. To the running of this website, we need your help to support us.

Thanks to everyone for your continued support.

No, Thanks

We need your sign to support Project to invent "SMART AND CONTROLLABLE REFLECTIVE BALLOONS" to cover the Sun and Save Our Earth.

More details...

Sign Now!

We are very appreciated for your Prompt Action!