Protein "fingerprinting" in complex mixtures with peptoid microarrays.

We report here that microarrays comprised of several thousand peptoids (oligo-N-substituted glycines) are useful tools for the identification of proteins via a "fingerprinting" approach. By using maltose-binding protein, glutathione S-transferase, and ubiquitin, a specific and highly reproducible pattern of binding was observed when fluorescently labeled protein was hybridized to the array. A similar pattern was obtained when binding of an unlabeled protein to the array was visualized by secondary hybridization of a labeled antibody against that protein, showing that native proteins can be identified without the requirement for prior chemical labeling. This work suggests that small-molecule microarrays might be used for more complex fingerprinting assays of potential diagnostic value.

[1]  Hanspeter Herzel,et al.  Identification of distinct antibody epitopes and mimotopes from a peptide array of 5520 randomly generated sequences. , 2002, Journal of immunological methods.

[2]  Stephen B. H. Kent,et al.  Efficient method for the preparation of peptoids [oligo(N-substituted glycines)] by submonomer solid-phase synthesis , 1992 .

[3]  T. Kodadek,et al.  Microwave-assisted solid-phase synthesis of peptoids. , 2002, Organic letters.

[4]  Stuart L. Schreiber,et al.  Printing Small Molecules as Microarrays and Detecting Protein−Ligand Interactions en Masse , 1999 .

[5]  R. Zuckermann,et al.  Synthesis of N-substituted glycine peptoid libraries. , 1996, Methods in enzymology.

[6]  S. P. Fodor,et al.  Light-directed, spatially addressable parallel chemical synthesis. , 1991, Science.

[7]  Jens Schneider-Mergener,et al.  Synthesis and screening of peptoid arrays on cellulose membranes , 2003 .

[8]  E. Petricoin,et al.  Rapid protein display profiling of cancer progression directly from human tissue using a protein biochip , 2000 .

[9]  E. Petricoin,et al.  Early detection: Proteomic applications for the early detection of cancer , 2003, Nature Reviews Cancer.

[10]  P. Clemons,et al.  A one-bead, one-stock solution approach to chemical genetics: part 2. , 2001, Chemistry & biology.

[11]  Laura Baldini,et al.  Pattern-based detection of different proteins using an array of fluorescent protein surface receptors. , 2004, Journal of the American Chemical Society.

[12]  R. Axel,et al.  The molecular biology of olfactory perception. , 1996, Cold Spring Harbor symposia on quantitative biology.

[13]  Kiyoshi Nokihara,et al.  Construction of a protein-detection system using a loop peptide library with a fluorescence label. , 2003, Chemistry & biology.

[14]  Kenji Usui,et al.  Peptide arrays with designed secondary structures for protein characterization using fluorescent fingerprint patterns. , 2004, Biopolymers.

[15]  Virginia Espina,et al.  Clinical proteomics: revolutionizing disease detection and patient tailoring therapy. , 2004, Journal of proteome research.

[16]  Jennifer L. Harris,et al.  Substrate profiling of deubiquitin hydrolases with a positional scanning library and mass spectrometry. , 2004, Biochemistry.

[17]  Thomas Kodadek,et al.  Isolation of protein ligands from large peptoid libraries. , 2003, Journal of the American Chemical Society.

[18]  J B Shear,et al.  Development of multianalyte sensor arrays composed of chemically derivatized polymeric microspheres localized in micromachined cavities. , 2001, Journal of the American Chemical Society.

[19]  Kit S Lam,et al.  From combinatorial chemistry to chemical microarray. , 2002, Current opinion in chemical biology.

[20]  J. Shand,et al.  An immobilised peptide array identifies antibodies to a discontinuous epitope in the extracellular domain of the bovine growth hormone receptor. , 1996, European journal of biochemistry.

[21]  R. Frank The SPOT-synthesis technique. Synthetic peptide arrays on membrane supports--principles and applications. , 2002, Journal of immunological methods.

[22]  Neal A. Rakow,et al.  A colorimetric sensor array for odour visualization , 2000, Nature.

[23]  T. Kodadek,et al.  Transformation of low-affinity lead compounds into high-affinity protein capture agents. , 2004, Chemistry & biology.

[24]  T. Cradick,et al.  Defining critical residues in the epitope for a HIV-neutralizing monoclonal antibody using phage display and peptide array technologies. , 1993, Gene.

[25]  T. Kodadek,et al.  Photolithographic synthesis of peptoids. , 2004, Journal of the American Chemical Society.

[26]  John A. Tallarico,et al.  A one-bead, one-stock solution approach to chemical genetics: part 1. , 2001, Chemistry & biology.

[27]  Stuart L Schreiber,et al.  Discovery of an inhibitor of a transcription factor using small molecule microarrays and diversity-oriented synthesis. , 2003, Journal of the American Chemical Society.

[28]  K. Lam,et al.  Peptide and small molecule microarray for high throughput cell adhesion and functional assays. , 2001, Bioconjugate chemistry.

[29]  M. Lesaicherre,et al.  Antibody-based fluorescence detection of kinase activity on a peptide array. , 2002, Bioorganic & medicinal chemistry letters.

[30]  Mahesh Uttamchandani,et al.  Small molecule microarrays: recent advances and applications. , 2005, Current opinion in chemical biology.

[31]  B. Cravatt,et al.  Discovering disease-associated enzymes by proteome reactivity profiling. , 2004, Chemistry & biology.

[32]  P. Schultz,et al.  Profiling protein function with small molecule microarrays , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[33]  M. Rechsteiner,et al.  Substrate specificity of the human proteasome. , 2001, Chemistry & biology.

[34]  Stuart L. Schreiber,et al.  Dissecting glucose signalling with diversity-oriented synthesis and small-molecule microarrays , 2002, Nature.

[35]  L Wang,et al.  Peptoids: a modular approach to drug discovery. , 1992, Proceedings of the National Academy of Sciences of the United States of America.