Periplasmically-exported lupanine hydroxylase undergoes transition from soluble to functional inclusion bodies in Escherichia coli

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dc.contributor.author Stampolidis, P.
dc.contributor.author Kaderbhai, Naheed N.
dc.contributor.author Kaderbhai, Mustak A.
dc.date.accessioned 2010-03-17T09:50:54Z
dc.date.available 2010-03-17T09:50:54Z
dc.date.issued 2009-01-23
dc.identifier.citation Stampolidis , P , Kaderbhai , N N & Kaderbhai , M A 2009 , ' Periplasmically-exported lupanine hydroxylase undergoes transition from soluble to functional inclusion bodies in Escherichia coli ' Archives of Biochemistry and Biophysics , vol 484 , no. 1 , pp. 8-15 . , 10.1016/j.abb.2009.01.017 en
dc.identifier.issn 1096-0384
dc.identifier.other PURE: 145049
dc.identifier.other dspace: 2160/4454
dc.identifier.uri http://hdl.handle.net/2160/4454
dc.description Stampolidis, P., Kaderbhai, N. N., Kaderbhai, M. A. (2009). Periplasmically-exported lupanine hydroxylase undergoes transition from soluble to functional inclusion bodies in Escherichia coli. Archives of Biochemistry and Biophysics, 484, (1), 8-15. On file IMPF: 03.05 RONO: 00 en
dc.description.abstract Pseudomonas lupanine hydroxylase is a periplasmic-localised, two domain quinocytochrome c enzyme. It requires numerous post-translocation modifications involving signal peptide processing, disulphide bridge formation and, heme linkage in the carboxy-terminal cytochrome c domain to eventually generate a Ca2+-bound quino-c hemoprotein that hydroxylates the plant alkaloid, lupanine. An exported, functional recombinant enzyme was generated in Escherichia coli by co-expression with cytochrome c maturation factors. Increased growth temperatures ranging from 18 to 30 °C gradually raised the enzyme production to a peak together with its concomitant aggregation as red solid particles, readily activatable in a fully functional form by mild chaotropic treatment. Here, we demonstrate that the exported lupanine hydroxylase undergoes a cascade transition from a soluble to “non-classical” inclusion body form when build-up in the periplasm exceeded a basal threshold concentration. These periplasmic aggregates were distinct from the non-secreted, signal-sequenceless counterpart that occurred as misfolded, non-functional concatamers in the form of classical inclusion bodies. We discuss our findings in the light of current models of how aggregation of lupanine hydroxylase arises in the periplasmic space. en
dc.format.extent 8 en
dc.language.iso eng
dc.relation.ispartof Archives of Biochemistry and Biophysics en
dc.subject Protein export en
dc.subject Periplasmic space en
dc.subject Amorphous protein aggregation en
dc.subject LH en
dc.subject Inclusion bodies en
dc.subject Disulphide-bond formation en
dc.subject Pyrroloquinoline quinine en
dc.subject Post-translational modifications en
dc.subject Quinocytochrome c en
dc.title Periplasmically-exported lupanine hydroxylase undergoes transition from soluble to functional inclusion bodies in Escherichia coli en
dc.type Text en
dc.type.publicationtype Article (Journal) en
dc.identifier.doi http://dx.doi.org/10.1016/j.abb.2009.01.017
dc.contributor.institution Institute of Biological, Environmental and Rural Sciences en
dc.description.status Peer reviewed en


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