Probing the Assembly of HDL Mimetic, Drug Carrying Nanoparticles Using Intrinsic Fluorescence

Raut, Sangram; Garud, Ashwini; Nagarajan, Bhavanai; Sabnis, Nirupama; Remaley, Alan; Fudala, Rafal; Gryczynski, Ignacy; Gryczynski, Zygmunt; Dzyuba, Sergei V.; Borejdo, Julian; Lacko, Andras

Publication

Probing the Assembly of HDL Mimetic, Drug Carrying Nanoparticles Using Intrinsic Fluorescence

Raut, Sangram; Garud, Ashwini; Nagarajan, Bhavanai; Sabnis, Nirupama; Remaley, Alan; Fudala, Rafal; Gryczynski, Ignacy; Gryczynski, Zygmunt; Dzyuba, Sergei V.; Borejdo, Julian; Lacko, Andras

Publisher

ASPET

Date

2020-03-16

Abstract

Reconstituted high-density lipoprotein (HDL) containing apolipoprotein A-I (Apo A-I) mimics the structure and function of endogenous (human plasma) HDL due to its function and potential therapeutic utility in atherosclerosis, cancer, neurodegenerative diseases, and inflammatory diseases. Recently, a new class of HDL mimetics has emerged, involving peptides with amino acid sequences that simulate the the primary structure of the amphipathic alpha helices within the Apo A-I protein. The findings reported in this communication were obtained using a similar amphiphilic peptide (modified via conjugation of a myristic acid residue at the amino terminal aspartic acid) that self-assembles (by itself) into nanoparticles while retaining the key features of endogenous HDL. The studies presented here involve the macromolecular assembly of the myristic acid conjugated peptide (MYR-5A) into nanomicellar structures and its characterization via steady-state and time-resolved fluorescence spectroscopy. The structural differences between the free peptide (5A) and MYR-5A conjugate were also probed, using tryptophan fluorescence, Förster resonance energy transfer (FRET), dynamic light scattering, and gel exclusion chromatography. To our knowledge, this is the first report of a lipoprotein assembly generated from a single ingredient and without a separate lipid component. The therapeutic utility of these nanoparticles (due to their capablity to incorporate a wide range of drugs into their core region for targeted delivery) was also investigated by probing the role of the scavenger receptor type B1 in this process.