Export of a hyper-expressed mammalian globular cytochrome b5 precursor in Escherichia coli is dramatically affected by the nature of the amino acid flanking the secretory signal sequence cleavage bond

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dc.contributor.author Kaderbhai, Naheed N.
dc.contributor.author Ahmed, Khalil
dc.contributor.author Kaderbhai, Mustak A.
dc.date.accessioned 2011-06-01T07:52:39Z
dc.date.available 2011-06-01T07:52:39Z
dc.date.issued 2010-07
dc.identifier.citation Kaderbhai , N N , Ahmed , K & Kaderbhai , M A 2010 , ' Export of a hyper-expressed mammalian globular cytochrome b5 precursor in Escherichia coli is dramatically affected by the nature of the amino acid flanking the secretory signal sequence cleavage bond ' Protein Science , vol 19 , no. 7 , pp. 1344-1353 . en
dc.identifier.issn 1469-896X
dc.identifier.other PURE: 162721
dc.identifier.other dspace: 2160/6850
dc.identifier.uri http://hdl.handle.net/2160/6850
dc.description Kaderbhai, N.N., Ahmed, K., Kaderbhai, M.A. (2010). Export of a hyper-expressed mammalian globular cytochrome b5 precursor in Escherichia coli is dramatically affected by the nature of the amino acid flanking the secretory signal sequence cleavage bond. Protein Science, 19 (7), 1344-1353. IMPF: 02.74 en
dc.description.abstract A chimeric mammalian globular cytochrome b(5) fused to Escherichia coil alkaline phosphatase signal sequence (SS) was used as a model probe to investigate the influence of substituting each one of the standard 20 amino acids at its N-terminus on the Sec-dependent export of the precursor to the periplasmic space of E. coil. Substituting the native Met(+1) of the passenger protein flanking the SS with any one of the remaining 19 amino acids introduced significant changes in the export of cytochrome b(5) without jamming the Sec-dependent translocon. Acidic and hydrophilic residues proved to be the most efficient promoters of export. Small, nonbulky and basic residues yielded intermediate levels of the hemoprotein export. Replacement with a Cys(+1) residue generated significant quantities of both monomeric and disulfide-linked dimeric forms. However, bulky, aromatic and hydrophobic residues caused a significant decline in the rates of secretion. In expectation with their absences in the natural periplasmically secreted proteins, Pro and Ile-tagged cytochrome b(5) precursors failed to generate any detectable secreted recombinant products. Although Ala, amongst the native E. coil periplasmic proteins, is the preferred X+1 residue with an occurrence of 50% frequency, it proved half as effective in promoting export when inserted proximally to the SS of cytochrome b(5). The mechanisms involved for these export variations are discussed. The findings will prove beneficial for high-level generation of recombinant proteins by secretory means for pharmaceutical and related biotechnological applications. en
dc.format.extent 10 en
dc.language.iso eng
dc.relation.ispartof Protein Science en
dc.subject protein export en
dc.subject recombinant protein en
dc.subject cytochrome b5 en
dc.subject signal sequence en
dc.subject preprotein processing en
dc.subject periplasmic space en
dc.subject Escherichia coli en
dc.title Export of a hyper-expressed mammalian globular cytochrome b5 precursor in Escherichia coli is dramatically affected by the nature of the amino acid flanking the secretory signal sequence cleavage bond en
dc.type Text en
dc.type.publicationtype Article (Journal) en
dc.identifier.doi http://dx.doi.org/10.1002/pro.411
dc.contributor.institution Institute of Biological, Environmental and Rural Sciences en
dc.description.status Peer reviewed en


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