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Tuning Liposome Membrane Permeability by Competitive Peptide Dimerization and Partitioning-Folding Interactions Regulated by Proteolytic Activity.

Lim SK, Sandén C, Selegård R, Liedberg B, Aili D - Sci Rep (2016)

Bottom Line: We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation.The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors.This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo.

View Article: PubMed Central - PubMed

Affiliation: Centre for Biomimetic Sensor Science, School of Materials Science and Engineering, Nanyang Technological University, Research Techno Plaza, 6th storey XFrontiers block, 50 Nanyang Drive, 637553 Singapore.

ABSTRACT
Membrane active peptides are of large interest for development of drug delivery vehicles and therapeutics for treatment of multiple drug resistant infections. Lack of specificity can be detrimental and finding routes to tune specificity and activity of membrane active peptides is vital for improving their therapeutic efficacy and minimize harmful side effects. We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation. Dimerization with a complementary peptide efficiently inhibits formation of pores. The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors. This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo.

No MeSH data available.


Related in: MedlinePlus

POPC liposomes with varying amounts (1–10 mol%) of a maleimide head group functionalized lipid (MPB-PE) were prepared (top).The MPB-PE lipid anchored the de novo designed helix-loop-helix polypeptide JR2KC to the membrane, which triggered pore formation (left). The pore formation was inhibited by addition of the charge complementary polypeptide JR2E that heterodimerize with JR2KC and folds into a four-helix bundle (right). Not drawn to scale.
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f1: POPC liposomes with varying amounts (1–10 mol%) of a maleimide head group functionalized lipid (MPB-PE) were prepared (top).The MPB-PE lipid anchored the de novo designed helix-loop-helix polypeptide JR2KC to the membrane, which triggered pore formation (left). The pore formation was inhibited by addition of the charge complementary polypeptide JR2E that heterodimerize with JR2KC and folds into a four-helix bundle (right). Not drawn to scale.

Mentions: Here, we describe a de novo designed membrane active helix-loop-helix peptide (JR2KC) that partition into zwitterionic lipid membranes when specifically and covalently anchored to head-group functionalized lipids present in the membrane. Anchoring and subsequent membrane partitioning triggers a structural transition of the peptide from random coil to α-helical conformation and pore formation as schematically outlined in Fig. 1. The extent of pore formation can be dynamically tuned by varying the number of anchoring moieties or by introducing a complementary helix-loop-helix peptide (JR2E) that is designed to heterodimerize with JR2KC and fold into four-helix bundles21. Moreover, we show that the inhibiting activity of JR2E can be controlled enzymatically by the matrix metalloproteinase MMP-7. Upregulation of MMP-7 is associated with the progression of a range of malignant tumours and is thought to be involved in cancer metastasis and various inflammatory processes22. JR2E contains two specific recognition sites for MMP-7, at positions 11-Ala-/-Leu-12 and 26-Ala-/ Gln-27, and proteolytic cleavage of JR2E results in a truncated peptide23, that is unable to heterodimerize with JR2KC. MMP-7 thus reactivates the membrane permeabilizing effect of JR2KC. Here, we thus show a novel recognition based strategy for specific tuning of the permeability of lipid membranes using a de novo designed membrane active peptide. In addition to demonstrating a possible path for obtaining more specific designer AMPs, this system enables enzyme mediated liposomal release. The latter indicates a possible strategy for design of peptides for increasing efficacy of liposome-encapsulated anti-cancer drugs by increasing the release rate in the vicinity of tumors.


Tuning Liposome Membrane Permeability by Competitive Peptide Dimerization and Partitioning-Folding Interactions Regulated by Proteolytic Activity.

Lim SK, Sandén C, Selegård R, Liedberg B, Aili D - Sci Rep (2016)

POPC liposomes with varying amounts (1–10 mol%) of a maleimide head group functionalized lipid (MPB-PE) were prepared (top).The MPB-PE lipid anchored the de novo designed helix-loop-helix polypeptide JR2KC to the membrane, which triggered pore formation (left). The pore formation was inhibited by addition of the charge complementary polypeptide JR2E that heterodimerize with JR2KC and folds into a four-helix bundle (right). Not drawn to scale.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4759693&req=5

f1: POPC liposomes with varying amounts (1–10 mol%) of a maleimide head group functionalized lipid (MPB-PE) were prepared (top).The MPB-PE lipid anchored the de novo designed helix-loop-helix polypeptide JR2KC to the membrane, which triggered pore formation (left). The pore formation was inhibited by addition of the charge complementary polypeptide JR2E that heterodimerize with JR2KC and folds into a four-helix bundle (right). Not drawn to scale.
Mentions: Here, we describe a de novo designed membrane active helix-loop-helix peptide (JR2KC) that partition into zwitterionic lipid membranes when specifically and covalently anchored to head-group functionalized lipids present in the membrane. Anchoring and subsequent membrane partitioning triggers a structural transition of the peptide from random coil to α-helical conformation and pore formation as schematically outlined in Fig. 1. The extent of pore formation can be dynamically tuned by varying the number of anchoring moieties or by introducing a complementary helix-loop-helix peptide (JR2E) that is designed to heterodimerize with JR2KC and fold into four-helix bundles21. Moreover, we show that the inhibiting activity of JR2E can be controlled enzymatically by the matrix metalloproteinase MMP-7. Upregulation of MMP-7 is associated with the progression of a range of malignant tumours and is thought to be involved in cancer metastasis and various inflammatory processes22. JR2E contains two specific recognition sites for MMP-7, at positions 11-Ala-/-Leu-12 and 26-Ala-/ Gln-27, and proteolytic cleavage of JR2E results in a truncated peptide23, that is unable to heterodimerize with JR2KC. MMP-7 thus reactivates the membrane permeabilizing effect of JR2KC. Here, we thus show a novel recognition based strategy for specific tuning of the permeability of lipid membranes using a de novo designed membrane active peptide. In addition to demonstrating a possible path for obtaining more specific designer AMPs, this system enables enzyme mediated liposomal release. The latter indicates a possible strategy for design of peptides for increasing efficacy of liposome-encapsulated anti-cancer drugs by increasing the release rate in the vicinity of tumors.

Bottom Line: We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation.The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors.This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo.

View Article: PubMed Central - PubMed

Affiliation: Centre for Biomimetic Sensor Science, School of Materials Science and Engineering, Nanyang Technological University, Research Techno Plaza, 6th storey XFrontiers block, 50 Nanyang Drive, 637553 Singapore.

ABSTRACT
Membrane active peptides are of large interest for development of drug delivery vehicles and therapeutics for treatment of multiple drug resistant infections. Lack of specificity can be detrimental and finding routes to tune specificity and activity of membrane active peptides is vital for improving their therapeutic efficacy and minimize harmful side effects. We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation. Dimerization with a complementary peptide efficiently inhibits formation of pores. The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors. This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo.

No MeSH data available.


Related in: MedlinePlus