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Bombesin peptide conjugated gold nanocages internalize via clathrin mediated endocytosis

Dhananjay Suresh1, Ajit Zambre, Nripen Chanda

  • 1Departments of †Bioengineering, ‡Radiology, ¥Medicine and §Chemistry, ⊥University of Missouri Research Reactor, and #International Center for Nano/Micro Systems and Nanotechnology, University of Missouri , Columbia, Missouri 65211, United States.

Bioconjugate Chemistry
|July 15, 2014
PubMed

Insights

Targeted gold nanocages (AuNCs) conjugated with bombesin peptide are effectively internalized by cancer cells via clathrin-mediated endocytosis (CME). This study clarifies nanoparticle uptake mechanisms for enhanced drug delivery systems.

Area of Science:

  • Biomedical Engineering
  • Nanotechnology
  • Cell Biology

Background:

  • The cellular internalization mechanisms of receptor-targeted nanoparticles are not fully understood.
  • Peptide-conjugated nanoparticles offer potential for targeted drug delivery, but their cellular interactions require detailed investigation.

Purpose of the Study:

  • To elucidate the cellular internalization mechanism and intracellular trafficking of peptide-conjugated gold nanocages (AuNCs) in cancer cells.
  • To confirm the target specificity and functional robustness of peptide ligands on nanoparticle surfaces for drug delivery applications.

Main Methods:

  • Synthesis and characterization of bombesin (BBN) peptide-conjugated gold nanocages (AuNC-BBN).
  • Radiolabeled competitive cell binding assays to assess affinity for gastrin-releasing peptide receptors (GRPR).
  • Cell inhibitor studies and microscopy (optical and transmission electron microscopy - TEM) to investigate endocytic pathways.

Main Results:

  • AuNC-BBN demonstrated selective binding to GRPR on PC3 cancer cells.
  • Cellular uptake of AuNC-BBN was predominantly mediated by clathrin-mediated endocytosis (CME), with an up to 84% reduction in uptake observed in the presence of CME inhibitors.
  • TEM imaging confirmed characteristic clathrin-coated pits and lysosomal release of AuNCs, validating the CME pathway.

Conclusions:

  • Peptide ligands conjugated to gold nanocages retain their target specificity and functionality.
  • Clathrin-mediated endocytosis is the primary mechanism for the cellular internalization of AuNC-BBN.
  • These findings support the use of peptide-conjugated nanoparticles as robust intracellular vehicular delivery systems.

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