Exploring the surface charge on peptide-gold nanoparticle conjugates by force spectroscopy.
The conformation and charge exposure of peptides attached to colloidal gold nanoparticles (AuNPs) are critical for both the colloidal stability and for the recognition of biological targets in biomedical applications such as diagnostics and therapy. We prepared conjugates of AuNPs and three isomer peptides capable of recognizing toxic aggregates of the amyloid beta protein (Abeta) involved in Alzheimer's disease, namely, CLPFFD-CONH(2) (i0), CDLPFF-CONH(2) (i1), and CLPDFF-CONH(2) (i2), where D is the amino acid aspartic acid that is negatively charged at pH = 7.4. We then studied the effect of peptide sequence on the charge exposure through force spectroscopy measurements. The peptide-AuNPs conjugates were fixed on glass surfaces, and their interactions with peptide-functionalized tips were determined. Our results show a higher density of surface charge in the conjugates of the isomers i0 and i2 and a lower density in i1, which is due to the higher degree of functionalization in the first two compared with the third. However, the charge per molecule of the peptide is higher for i1 with respect to i0 and i2, which could be related to the local conformation that the peptides adopt on the surface. The acid-base behavior of the peptide anchored to the AuNPs is different than expected in aqueous solutions of free peptides, which could be related to the low accessibility of the NH(2)-terminal group belonging to the cysteine that is located near the AuNPs surface. In contrast with other techniques, the fixation of the peptide-AuNPs conjugates to a surface allows for characterization of the local charge exposure of peptides anchored to AuNPs over a wide range of pH.
Guerrero
Ariel R
AR
Materials and Surface Science Group, University of Windsor, 401 Sunset Avenue, Windsor, Ontario N9B 3P4, Canada.
Caballero
Leonardo
L
Adeva
Alberto
A
Melo
Francisco
F
Kogan
Marcelo J
MJ
D000577:Amides / Q000737:chemistry; D000595:Amino Acid Sequence; D006046:Gold / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D007536:Isomerism; D053768:Metal Nanoparticles / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D008968:Molecular Conformation; D010455:Peptides / Q000737:chemistry*; D013057:Spectrum Analysis / Q000379:methods*; D013499:Surface Properties
D000577:Amides; D010455:Peptides; D006046:Gold