1 Ocak 2017 Dergi makalesi Açık Erişim

Mamedov, Tarlan; Cicek, Kader; Gulec, Burcu; Ungor, Rifat; Hasanova, Gulnara

DataCite XML

<?xml version='1.0' encoding='utf-8'?>
<resource xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns="http://datacite.org/schema/kernel-4" xsi:schemaLocation="http://datacite.org/schema/kernel-4 http://schema.datacite.org/meta/kernel-4.1/metadata.xsd">
  <identifier identifierType="URL">https://aperta.ulakbim.gov.tr/record/48931</identifier>
  <creators>
    <creator>
      <creatorName>Mamedov, Tarlan</creatorName>
      <givenName>Tarlan</givenName>
      <familyName>Mamedov</familyName>
      <affiliation>Akdeniz Univ, Dept Agr Biotechnol, Antalya, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Cicek, Kader</creatorName>
      <givenName>Kader</givenName>
      <familyName>Cicek</familyName>
      <affiliation>Akdeniz Univ, Dept Agr Biotechnol, Antalya, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Gulec, Burcu</creatorName>
      <givenName>Burcu</givenName>
      <familyName>Gulec</familyName>
      <affiliation>Akdeniz Univ, Dept Agr Biotechnol, Antalya, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Ungor, Rifat</creatorName>
      <givenName>Rifat</givenName>
      <familyName>Ungor</familyName>
      <affiliation>Akdeniz Univ, Dept Agr Biotechnol, Antalya, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Hasanova, Gulnara</creatorName>
      <givenName>Gulnara</givenName>
      <familyName>Hasanova</familyName>
      <affiliation>Akdeniz Univ, Dept Agr Biotechnol, Antalya, Turkey</affiliation>
    </creator>
  </creators>
  <titles>
    <title>In Vivo Production Of Non-Glycosylated Recombinant Proteins In Nicotiana Benthamiana Plants By Co-Expression With Endo-Beta-N-Acetylglucosaminidase H (Endo H) Of Streptomyces Plicatus</title>
  </titles>
  <publisher>Aperta</publisher>
  <publicationYear>2017</publicationYear>
  <dates>
    <date dateType="Issued">2017-01-01</date>
  </dates>
  <resourceType resourceTypeGeneral="Text">Journal article</resourceType>
  <alternateIdentifiers>
    <alternateIdentifier alternateIdentifierType="url">https://aperta.ulakbim.gov.tr/record/48931</alternateIdentifier>
  </alternateIdentifiers>
  <relatedIdentifiers>
    <relatedIdentifier relatedIdentifierType="DOI" relationType="IsIdenticalTo">10.1371/journal.pone.0183589</relatedIdentifier>
  </relatedIdentifiers>
  <rightsList>
    <rights rightsURI="http://www.opendefinition.org/licenses/cc-by">Creative Commons Attribution</rights>
    <rights rightsURI="info:eu-repo/semantics/openAccess">Open Access</rights>
  </rightsList>
  <descriptions>
    <description descriptionType="Abstract">A plant transient expression system, with eukaryotic post-translational modification machinery, offers superior efficiency, scalability, safety, and lower cost over other expression systems. However, due to aberrant N-glycosylation, this expression system may not be a suitable expression platform for proteins not carrying N-linked glycans in the native hosts. Therefore, it is crucial to develop a strategy to produce target proteins in a non-glycosylated form while preserving their native sequence, conformation and biological activity. Previously, we developed a strategy for enzymatic deglycosylation of proteins in planta by co-expressing bacterial peptide-N-glycosidase F (PNGase F). Though PNGase F removes oligosaccharides from glycosylated proteins, in so doing it causes an amino acid change due to the deamidation of asparagine to aspartate in the N-X-S/T site. Endo-beta-N-acetylglucosaminidase (EC3.2.1.96, Endo H), another deglycosylating enzyme, catalyzes cleavage between two N-Acetyl-D-glucosamine residues of the chitobiose core of N-linked glycans, leaving a single N-Acetyl-D-glucosamine residue without the concomitant deamidation of asparagine. In this study, a method for in vivo deglycosylation of recombinant proteins in plants by transient co-expression with bacterial Endo H is described for the first time. Endo H was fully active in vivo. and successfully cleaved N-linked glycans from glycoproteins were tested. In addition, unlike the glycosylated form, in vivo Endo H deglycosylated Pfs48/45 was recognized by conformational specific Pfs48/45 monoclonal antibody, in a manner similar to its PNGase F deglycosylated counterpart. Furthermore, the deglycosylated PA83 molecule produced by Endo H showed better stability than a PNGase F deglycosylated counterpart. Thus, an Endo H in vivo deglycosylation approach provides another opportunity to develop vaccine antigens, therapeutic proteins, antibodies, and industrial enzymes.</description>
  </descriptions>
</resource>