title: Exploration of possible binding sites of nanoparticles on protein by cross-linking chemistry coupled with mass spectrometry.
abstract: For the first time, the possible binding site of nanoparticles on protein was revealed by cross-linking chemistry coupled with mass spectrometry. The peptides located very close to the poly(acrylic acid) (PAA)-coated Fe(3)O(4) nanoparticles (NPs) during interaction with human serum albumin (HSA) were cross-linked to the surface of NPs. Following protease digestion, the attached peptides were cleaved off the particle surface and identified by matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS). The peptides were found to be part of the so-called drug binding site 2 of HSA; and the competitive binding to HSA between the corresponding drug, ibuprofen, and the NPs was observed. Our results demonstrated that cross-linking chemistry coupled with MS was a quick and simple method for locating the possible binding sites of NPs on protein. Information on NP-protein binding interface will benefit the study of how the interactions are governed by the physicochemical properties of NPs, for guiding the design of functional bionano constructs. It can also help to predict the biological consequence of protein adsorption on NPs, for obtaining more knowledge on nanotoxicity.
authors.forename: Ni
authors.initials: N
authors.lastname: Li
authors.forename: Shang
authors.initials: S
authors.lastname: Zeng
authors.forename: Le
authors.initials: L
authors.lastname: He
authors.forename: Wenwan
authors.initials: W
authors.lastname: Zhong
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D000595:Amino Acid Sequence; D001665:Binding Sites; D003432:Cross-Linking Reagents / Q000737:chemistry*; D052203:Ferrosoferric Oxide / Q000737:chemistry; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
chemical_list: D000180:Acrylic Resins; D003432:Cross-Linking Reagents; D010455:Peptides; D012709:Serum Albumin; C006903:carbopol 940; D014357:Trypsin; D052203:Ferrosoferric Oxide