Conformational and membrane-binding properties of a signal sequence are largely unaltered by its adjacent mature region.

McKnight, C. James; Stradley, Sarah J.; Jones, Jeffrey; Gierasch, Lila M.

doi:10.1073/pnas.88.13.5799

Cited by 31 publications

(20 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…23 Intriguingly, mutations in the early region of a passenger protein can abolish correct targeting, implying that there are restrictions on the allowed sequences of secreted proteins (e.g., Ref. 24).…”

Section: Signal Peptide Conformations and Membrane Interactionsmentioning

confidence: 99%

Use of synthetic signal sequences to explore the protein export machinery

Clérico¹,

Maki²,

Gierasch³

2008

Biopolymers

View full text Add to dashboard Cite

The information for correct localization of newly synthesized proteins in both prokaryotes and eukaryotes resides in self‐contained, often transportable targeting sequences. Of these, signal sequences specify that a protein should be secreted from a cell or incorporated into the cytoplasmic membrane. A central puzzle is presented by the lack of primary structural homology among signal sequences, although they share common features in their sequences. Synthetic signal peptides have enabled a wide range of studies of how these “zipcodes” for protein secretion are decoded and used to target proteins to the protein machinery that facilitates their translocation across and integration into membranes. We review research on how the information in signal sequences enables their passenger proteins to be correctly and efficiently localized. Synthetic signal peptides have made possible binding and crosslinking studies to explore how selectivity is achieved in recognition by the signal sequence‐binding receptors, signal recognition particle, or SRP, which functions in all organisms, and SecA, which functions in prokaryotes and some organelles of prokaryotic origins. While progress has been made, the absence of atomic resolution structures for complexes of signal peptides and their receptors has definitely left many questions to be answered in the future. © 2007 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 90: 307–319, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

show abstract

Section: Signal Peptide Conformations and Membrane Interactionsmentioning

confidence: 99%

Use of synthetic signal sequences to explore the protein export machinery

Clérico¹,

Maki²,

Gierasch³

2008

Biopolymers

View full text Add to dashboard Cite

show abstract

“…Apparently, binding of the precursor to liposomes is mediated primarily by interactions with the presequence peptide. Although this is the first study comparing the lipid binding properties of an authentic mitochondrial precursor protein and its presequence peptide, similar studies using a secretory bacterial protein (42,50) also concluded that the mature portion of the precursor protein did not have any effect on the binding of the signal peptide to lipids. In our case, the only difference detected between the presequence peptide and the precursor was on the kinetics of carboxyfluorescein release from liposomes (Fig.…”

Section: Discussionmentioning

confidence: 99%

Interaction of the Precursor to Mitochondrial Aspartate Aminotransferase and Its Presequence Peptide with Model Membranes

Doñate¹,

Yáñez²,

Iriarte

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

The possible contribution of the mature portion of a mitochondrial precursor protein to its interaction with membrane lipids is unclear. To address this issue, we examined the interaction of the precursor to mitochondrial aspartate aminotransferase (pmAAT) and of a synthetic peptide corresponding to the 29-residue presequence peptide (mAAT-pp) with anionic phospholipid vesicles. The affinity of mAAT-pp and pmAAT for anionic vesicles is nearly identical. Results obtained by analyzing the effect of mAAT-pp or full-length pmAAT on either the permeability or microviscosity of the phospholipid vesicles are consistent with only a shallow insertion of the presequence peptide in the bilayer. Analysis of the quenching of Trp-17 fluorescence by brominated phospholipids reveals that this presequence residue inserts to a depth of approximately 9 Å from the center of the bilayer. Furthermore, in membrane-bound pmAAT or mAAT-pp, both Arg-8 and Arg-28 are accessible to the solvent. These results suggest that the presequence segment lies close to the surface of the membrane and that the mature portion of the precursor protein has little effect on the affinity or mode of binding of the presequence to model membranes. In the presence of vesicles, mAAT-pp adopts considerable ␣-helical structure. Hydrolysis by trypsin after Arg-8 results in the dissociation of the remaining 21-residue C-terminal peptide fragment from the membrane bilayer, suggesting that the N-terminal portion of the presequence is essential for membrane binding. Based on these results, we propose that the presequence peptide may contain dual recognition elements for both the lipid and import receptor components of the mitochondrial membrane.

show abstract

“…Many of these were conducted with constructs containing motifs around the cleavage site that differed markedly in their primary structure, appended to distinct passenger molecules. 1,23 A mammalian globular cytochrome b 5 (Cyt) tagged with(out) the alkaline phosphatase SS of E. coli has been exploited to investigate many diverse molecular features of high-level periplasmic protein secretion. 18,[24][25][26][27][28] It has proved an invaluable tool in reporting both the in vitro and in vivo sec-dependent status of recombinant protein generation by visual and spectroscopic means.…”

Section: Introductionmentioning

confidence: 99%

Export of a hyperexpressed 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

2010

View full text Add to dashboard Cite

Export of a hyperexpressed mammalian globular cytochrome b 5 precursor in Escherichia coli is dramatically affected by the nature of the amino acid flanking the secretory signal sequence cleavage bond Naheed N. Kaderbhai,* Khalil Ahmed, and Mustak A. Kaderbhai Abstract: A chimeric mammalian globular cytochrome b 5 fused to Escherichia coli 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. coli. Substituting the native Met 11 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 11 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. coli periplasmic proteins, is the preferred X 11 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.

show abstract

Conformational and membrane-binding properties of a signal sequence are largely unaltered by its adjacent mature region.

Cited by 31 publications

References 45 publications

Use of synthetic signal sequences to explore the protein export machinery

Use of synthetic signal sequences to explore the protein export machinery

Interaction of the Precursor to Mitochondrial Aspartate Aminotransferase and Its Presequence Peptide with Model Membranes

Export of a hyperexpressed 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

Contact Info

Product

Resources

About

Conformational and membrane-binding properties of a signal sequence are largely unaltered by its adjacent mature region.

Cited by 31 publications

References 45 publications

Use of synthetic signal sequences to explore the protein export machinery

Use of synthetic signal sequences to explore the protein export machinery

Interaction of the Precursor to Mitochondrial Aspartate Aminotransferase and Its Presequence Peptide with Model Membranes

Export of a hyperexpressed 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

Contact Info

Product

Resources

About

Export of a hyperexpressed 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