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Title: Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1

Abstract

The ability to orient biologically active proteins on surfaces is a major challenge in the design, construction, and successful deployment of many medical technologies. As methods to orient biomolecules are developed, it is also essential to develop techniques that can accurately determine the orientation and structure of these materials. In this study, two model protein and peptide systems are presented to highlight the strengths of three surface analysis techniques for characterizing protein films: time-of-flight secondary-ion mass spectrometry (ToF-SIMS), sum-frequency generation (SFG) vibrational spectroscopy, and near-edge x-ray absorption fine structure (NEXAFS) spectroscopy. First, the orientation of Protein G B1, a rigid 6 kDa domain covalently attached to a maleimide-functionalized self-assembled monolayer, was examined using ToF-SIMS. Although the thickness of the Protein G layer was similar to the ToF-SIMS sampling depth, orientation of Protein G was successfully determined by analyzing the C{sub 2}H{sub 5}S{sup +} intensity, a secondary-ion derived from a methionine residue located at one end of the protein. Next, the secondary structure of a 13-mer leucine-lysine peptide (LK{sub 310}) adsorbed onto hydrophilic quartz and hydrophobic fluorocarbon surfaces was examined. SFG spectra indicated that the peptide's lysine side chains were ordered on the quartz surface, while the peptide's leucine side chainsmore » were ordered on the fluorocarbon surface. NEXAFS results provided complementary information about the structure of the LK{sub 310} film and the orientations of amide bonds within the LK{sub 310} peptide.« less

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
Sponsoring Org.:
DOE - OFFICE OF SCIENCE
OSTI Identifier:
1019959
Report Number(s):
BNL-95805-2011-JA
Journal ID: ISSN 1071-1023; TRN: US201115%%595
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
Journal of Vacuum Science and Technology B: Microelectronics and Nanometer Structures
Additional Journal Information:
Journal Volume: 28; Journal Issue: 4; Journal ID: ISSN 1071-1023
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; ABSORPTION; AMIDES; CHAINS; CONSTRUCTION; DESIGN; FINE STRUCTURE; LEUCINE; LYSINE; MASS SPECTROSCOPY; METHIONINE; ORIENTATION; PEPTIDES; PROTEINS; QUARTZ; RESIDUES; SAMPLING; SPECTRA; SPECTROSCOPY; THICKNESS; national synchrotron light source

Citation Formats

Baio, J, Weidner, T, Samuel, N, McCrea, K, Baugh, L, Stayton, P, and Castner, D. Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1. United States: N. p., 2010. Web.
Baio, J, Weidner, T, Samuel, N, McCrea, K, Baugh, L, Stayton, P, & Castner, D. Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1. United States.
Baio, J, Weidner, T, Samuel, N, McCrea, K, Baugh, L, Stayton, P, and Castner, D. 2010. "Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1". United States.
@article{osti_1019959,
title = {Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1},
author = {Baio, J and Weidner, T and Samuel, N and McCrea, K and Baugh, L and Stayton, P and Castner, D},
abstractNote = {The ability to orient biologically active proteins on surfaces is a major challenge in the design, construction, and successful deployment of many medical technologies. As methods to orient biomolecules are developed, it is also essential to develop techniques that can accurately determine the orientation and structure of these materials. In this study, two model protein and peptide systems are presented to highlight the strengths of three surface analysis techniques for characterizing protein films: time-of-flight secondary-ion mass spectrometry (ToF-SIMS), sum-frequency generation (SFG) vibrational spectroscopy, and near-edge x-ray absorption fine structure (NEXAFS) spectroscopy. First, the orientation of Protein G B1, a rigid 6 kDa domain covalently attached to a maleimide-functionalized self-assembled monolayer, was examined using ToF-SIMS. Although the thickness of the Protein G layer was similar to the ToF-SIMS sampling depth, orientation of Protein G was successfully determined by analyzing the C{sub 2}H{sub 5}S{sup +} intensity, a secondary-ion derived from a methionine residue located at one end of the protein. Next, the secondary structure of a 13-mer leucine-lysine peptide (LK{sub 310}) adsorbed onto hydrophilic quartz and hydrophobic fluorocarbon surfaces was examined. SFG spectra indicated that the peptide's lysine side chains were ordered on the quartz surface, while the peptide's leucine side chains were ordered on the fluorocarbon surface. NEXAFS results provided complementary information about the structure of the LK{sub 310} film and the orientations of amide bonds within the LK{sub 310} peptide.},
doi = {},
url = {https://www.osti.gov/biblio/1019959}, journal = {Journal of Vacuum Science and Technology B: Microelectronics and Nanometer Structures},
issn = {1071-1023},
number = 4,
volume = 28,
place = {United States},
year = {Fri Jan 01 00:00:00 EST 2010},
month = {Fri Jan 01 00:00:00 EST 2010}
}