Interaction of peptides corresponding to mitochondrial presequences with membranes.

The transport of the F1-ATPase beta-subunit precursor into mitochondria is dependent upon a presequence at its amino terminus. Within the mitochondrial membrane translocation site the potential amphiphilic character of the presequence region may be necessary to stabilize binding to the mitochondrial inner membrane. To better understand its role in protein import, the interaction of the F1 beta-presequence with lipid membranes was measured using circular dichroism and surface tensiometry. These studies reveal that a 20-residue peptide containing the F1 beta-presequence binds to phospholipid vesicles (Kd = 4.5-6.0 x 10(-8)M and adopts a predominantly alpha-helical structure. Although the presequence peptide binds avidly to lipids, it does not appear to penetrate deeply into the bilayer to perturb a reporter probe in the membrane interior. Compared with the effect of the peptides with demonstrated membrane insertion and lytic properties, the F1-beta-presequence appears to displace phospholipid head groups but not insert deeply into the bilayer. High concentrations (greater than 50 microM) of presequence peptides are required to noticibly perturb import of the full length F1 alpha- or F1 beta-subunit precursors. Thus, the F1 beta-presequence alone is not sufficient to efficiently compete for import but may require a protein context or a minimal length to assist insertion into the transport site. These observations are discussed in light of the different requirements for import of various presequence containing precursors into mitochondria.

• Publication year

  1991

• Venue

  Journal of Biological Chemistry

• Publication date

  1991-11-15

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0021-9258(18)54692-5PMID 1834660
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• The F1 beta-presequence alone is not sufficient to efficiently compete for import and may require protein context or a minimal length to assist insertion into the transport site.

  Confidence 0.87

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• Presequence peptides at concentrations greater than 50 microM are required to noticeably perturb import of the full-length F1 alpha- or F1 beta-subunit precursors.

  Confidence 0.93

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• The F1 beta-presequence peptide does not appear to penetrate deeply into the bilayer or perturb a reporter probe in the membrane interior.

  Confidence 0.96

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• A 20-residue peptide containing the F1 beta-presequence binds to phospholipid vesicles with high affinity and adopts a predominantly alpha-helical structure.

  Confidence 0.98

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review

• alpha-helical structure

  secondary structure, structural state

  A protein secondary structure in which the peptide backbone forms a right-handed helix.

  Aliases: alpha helix, helical structure

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• bilayer interior reporter probe

  probe, measurement reagent

  A membrane-embedded probe used to sense perturbations in the interior region of a lipid bilayer.

  Aliases: membrane interior probe, reporter probe

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• f1 beta-presequence peptide

  peptide, mitochondrial targeting sequence

  A 20-residue amino-terminal peptide corresponding to the mitochondrial targeting presequence of the F1-ATPase beta-subunit.

  Aliases: F1 beta-presequence, beta-presequence peptide

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• full-length f1 alpha-subunit precursor

  protein precursor, import substrate

  The complete precursor form of the F1-ATPase alpha subunit that is imported into mitochondria.

  Aliases: F1 alpha-subunit precursor, alpha-subunit precursor

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• full-length f1 beta-subunit precursor

  protein precursor, import substrate

  The complete precursor form of the F1-ATPase beta subunit that is imported into mitochondria.

  Aliases: F1 beta-subunit precursor, beta-subunit precursor

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• minimal length

  sequence property, structural property

  The shortest peptide extent needed for a sequence to display the import-related behavior discussed here.

  Aliases: minimum length

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• mitochondrial inner membrane

  membrane, organelle membrane

  The inner lipid membrane of mitochondria that forms the barrier crossed during protein import.

  Aliases: inner membrane

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• mitochondrial transport site

  membrane site, transport site

  The membrane translocation region involved in moving precursor proteins across the mitochondrial inner membrane.

  Aliases: translocation site, import site

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• phospholipid vesicles

  model membrane, material

  Model membrane vesicles made of phospholipids that are used here to test peptide binding to lipid bilayers.

  Aliases: lipid vesicles, vesicles

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• presequence peptides

  peptide, experimental reagent

  Synthetic peptides derived from mitochondrial targeting presequences and used as import competitors in the membrane assay.

  Aliases: presequence peptide

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review
• protein context

  molecular context, property

  The surrounding polypeptide or precursor environment that can influence how a targeting sequence behaves during import.

  Aliases: context

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) review

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  1989cited by this paper
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  1989cited by this paper
• Mitochondrial protein import.

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• 70-kD heat shock-related protein is one of at least two distinct cytosolic factors stimulating protein import into mitochondria

  1988cited by this paper
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  1987cited by this paper
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