TY - JOUR
T1 - Quantitative Submonolayer Spatial Mapping of Arg-Gly-Asp-Containing Peptide Organomercaptan Gradients on Gold with Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry
AU - Wang, Qian
AU - Jakubowski, Jennifer A.
AU - Sweedler, Jonathan V.
AU - Bohn, Paul W.
PY - 2004/1/1
Y1 - 2004/1/1
N2 - Peptides containing the tripeptide sequence Arg-Gly-Asp (RGD) have the ability to bind to members of the integrin superfamily of cell-surface receptors and direct cellular adhesion and haptotaxis. The goal of this work is the development of a rapid and effective method for the quantitative submonolayer spatial composition mapping of surfaces displaying molecular assemblies of RGD-containing organomercaptan peptides on a Au surface using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALI)I-MS). Quantitation of the RGD peptide is achieved by determining the peak intensity of the protonated molecular ion, (M + H)+, relative to the (M + H)+ peak for an internal standard, which is similar chemically but with glutamic acid (E) substituted for aspartic acid (D). Using optimized sample preparation procedures, a bilinear calibration was obtained between the quantitative peak intensity ratio and the mole fraction of the RGD-containing peptide. Quantitative compositions were determined with relative standard deviations of < 10%, even in the presence of 10x spot-to-spot variations in the absolute signal intensities, by using this internal standard approach. This MALDI-MS quantitative analysis method was employed to probe variable-width two-component counterpropagating electrochemically generated gradients of the two peptides, prepared by coupling in-plane electrochemical potential gradients with the electrosorption reactions of organothiols to vary the composition laterally. The measured lateral composition profiles match the quasi-linear potential gradient model and yield profiles that overlap to a high degree of fidelity in potential space. Thus, MALDI-MS spatial composition mapping should become a powerful tool for the preparation of designed surfaces facilitating the study of cellular adhesion and motility and cell-cell interactions.
AB - Peptides containing the tripeptide sequence Arg-Gly-Asp (RGD) have the ability to bind to members of the integrin superfamily of cell-surface receptors and direct cellular adhesion and haptotaxis. The goal of this work is the development of a rapid and effective method for the quantitative submonolayer spatial composition mapping of surfaces displaying molecular assemblies of RGD-containing organomercaptan peptides on a Au surface using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALI)I-MS). Quantitation of the RGD peptide is achieved by determining the peak intensity of the protonated molecular ion, (M + H)+, relative to the (M + H)+ peak for an internal standard, which is similar chemically but with glutamic acid (E) substituted for aspartic acid (D). Using optimized sample preparation procedures, a bilinear calibration was obtained between the quantitative peak intensity ratio and the mole fraction of the RGD-containing peptide. Quantitative compositions were determined with relative standard deviations of < 10%, even in the presence of 10x spot-to-spot variations in the absolute signal intensities, by using this internal standard approach. This MALDI-MS quantitative analysis method was employed to probe variable-width two-component counterpropagating electrochemically generated gradients of the two peptides, prepared by coupling in-plane electrochemical potential gradients with the electrosorption reactions of organothiols to vary the composition laterally. The measured lateral composition profiles match the quasi-linear potential gradient model and yield profiles that overlap to a high degree of fidelity in potential space. Thus, MALDI-MS spatial composition mapping should become a powerful tool for the preparation of designed surfaces facilitating the study of cellular adhesion and motility and cell-cell interactions.
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U2 - 10.1021/ac030335+
DO - 10.1021/ac030335+
M3 - Article
C2 - 14697025
AN - SCOPUS:0346731183
SN - 0003-2700
VL - 76
SP - 1
EP - 8
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 1
ER -