SwePub
Sök i LIBRIS databas

  Utökad sökning

WFRF:(Villarejo Arsenio)
 

Sökning: WFRF:(Villarejo Arsenio) > (2011) > Importance of post-...

  • Burén, StefanUmeå universitet,Institutionen för fysiologisk botanik,Umeå Plant Science Centre (UPSC),Umea Univ, Dept Plant Physiol, Umea Plant Sci Ctr, S-90187 Umea, Sweden. (författare)

Importance of post-translational modifications for functionality of a chloroplast-localized carbonic anhydrase (CAH1) in Arabidopsis thaliana

  • Artikel/kapitelEngelska2011

Förlag, utgivningsår, omfång ...

  • 2011-06-10
  • Public Library of Science,2011
  • electronicrdacarrier

Nummerbeteckningar

  • LIBRIS-ID:oai:DiVA.org:umu-45474
  • https://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-45474URI
  • https://doi.org/10.1371/journal.pone.0021021DOI
  • https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-514132URI

Kompletterande språkuppgifter

  • Språk:engelska
  • Sammanfattning på:engelska

Ingår i deldatabas

Klassifikation

  • Ämneskategori:ref swepub-contenttype
  • Ämneskategori:art swepub-publicationtype

Anmärkningar

  • This work was supported by the Swedish Research Council (VR), the Kempe Foundations and Carl Tryggers Foundation to GS, and grant numbers BIO2006-08946 and BIO2009-11340 from the Spanish Ministerio de Ciencia e Innovacio´n (MICINN) to AV. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
  • BackgroundThe Arabidopsis CAH1 alpha-type carbonic anhydrase is one of the few plant proteins known to be targeted to the chloroplast through the secretory pathway. CAH1 is post-translationally modified at several residues by the attachment of N-glycans, resulting in a mature protein harbouring complex-type glycans. The reason of why trafficking through this non-canonical pathway is beneficial for certain chloroplast resident proteins is not yet known. Therefore, to elucidate the significance of glycosylation in trafficking and the effect of glycosylation on the stability and function of the protein, epitope-labelled wild type and mutated versions of CAH1 were expressed in plant cells.Methodology/Principal FindingsTransient expression of mutant CAH1 with disrupted glycosylation sites showed that the protein harbours four, or in certain cases five, N-glycans. While the wild type protein trafficked through the secretory pathway to the chloroplast, the non-glycosylated protein formed aggregates and associated with the ER chaperone BiP, indicating that glycosylation of CAH1 facilitates folding and ER-export. Using cysteine mutants we also assessed the role of disulphide bridge formation in the folding and stability of CAH1. We found that a disulphide bridge between cysteines at positions 27 and 191 in the mature protein was required for correct folding of the protein. Using a mass spectrometric approach we were able to measure the enzymatic activity of CAH1 protein. Under circumstances where protein N-glycosylation is blocked in vivo, the activity of CAH1 is completely inhibited.Conclusions/SignificanceWe show for the first time the importance of post-translational modifications such as N-glycosylation and intramolecular disulphide bridge formation in folding and trafficking of a protein from the secretory pathway to the chloroplast in higher plants. Requirements for these post-translational modifications for a fully functional native protein explain the need for an alternative route to the chloroplast.

Ämnesord och genrebeteckningar

Biuppslag (personer, institutioner, konferenser, titlar ...)

  • Ortega-Villasante, CristinaUniv Autonoma Madrid, Dept Biol, Madrid, Spain. (författare)
  • Blanco-Rivero, AmayaUniv Autonoma Madrid, Dept Biol, Madrid, Spain. (författare)
  • Martínez-Bernardini, AndreaUmeå universitet,Institutionen för fysiologisk botanik,Umeå Plant Science Centre (UPSC),Umea Univ, Dept Plant Physiol, Umea Plant Sci Ctr, S-90187 Umea, Sweden.;Univ Autonoma Madrid, Dept Biol, Madrid, Spain. (författare)
  • Shutova, Tatiana,1964-Umeå universitet,Umeå Plant Science Centre (UPSC),Institutionen för fysiologisk botanik,Umea Univ, Dept Plant Physiol, Umea Plant Sci Ctr, S-90187 Umea, Sweden.(Swepub:umu)tash0001 (författare)
  • Shevela, DmitriyUmeå universitet,Kemiska institutionen,Umea Univ, Dept Chem, S-90187 Umea, Sweden.(Swepub:umu)dmsh0004 (författare)
  • Messinger, JohannesUmeå universitet,Kemiska institutionen,Institutionen för fysiologisk botanik,Umeå Plant Science Centre (UPSC),Umea Univ, Dept Plant Physiol, Umea Plant Sci Ctr, S-90187 Umea, Sweden.;Umea Univ, Dept Chem, S-90187 Umea, Sweden.(Swepub:uu)johme269 (författare)
  • Bako, Laszlo,1959-Umeå universitet,Institutionen för fysiologisk botanik,Umeå Plant Science Centre (UPSC),Umea Univ, Dept Plant Physiol, Umea Plant Sci Ctr, S-90187 Umea, Sweden.(Swepub:umu)laba0001 (författare)
  • Villarejo, ArsenioUniv Autonoma Madrid, Dept Biol, Madrid, Spain. (författare)
  • Samuelsson, Göran,1951-Umeå universitet,Institutionen för fysiologisk botanik,Umeå Plant Science Centre (UPSC),Umea Univ, Dept Plant Physiol, Umea Plant Sci Ctr, S-90187 Umea, Sweden.(Swepub:umu)gosa0002 (författare)
  • Umeå universitetInstitutionen för fysiologisk botanik (creator_code:org_t)

Sammanhörande titlar

  • Ingår i:PLOS ONE: Public Library of Science6:6, s. e21021-1932-6203

Internetlänk

Hitta via bibliotek

  • PLOS ONE (Sök värdpublikationen i LIBRIS)

Till lärosätets databas

Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.

 
pil uppåt Stäng

Kopiera och spara länken för att återkomma till aktuell vy