HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: T300008-MCP200v1 T300008-MCP200v2 3/3/266    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bieber, A. L. Articles by Nelson, R. W. Search for Related Content PubMed PubMed Citation Articles by Bieber, A. L. Articles by Nelson, R. W. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 3:266-272, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Technology

Metal Ligand Affinity Pipettes and Bioreactive Alkaline Phosphatase Probes

Tools for Characterization of Phosphorylated Proteins and Peptides^*

Allan L. Bieber^,, Kemmons A. Tubbs and Randall W. Nelson^,¶

From Intrinsic Bioprobes, Inc., Tempe, AZ 85281; and the Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287-1604

An alkaline phosphatase-bioreactive probe, in which the enzyme is covalently bound to the mass spectrometry target, has been developed for studies of phosphoproteins. The bioreactive probe was used in combination with affinity capture and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to study hydrolysis of several phosphoproteins found in human saliva. Human salivary proteins were extracted from diluted human saliva with immobilized metal-affinity pipettes, which under defined conditions bound the phosphoproteins of interest preferentially over histatins. Phosphoproteins were eluted directly from the affinity pipettes to the bioreactive probe with diluted ammonium hydroxide, which provided conditions appropriate for hydrolysis by the alkaline phosphatase covalently bound to the probe surface. Results indicate the combination of metal-affinity pipette extraction, alkaline phosphatase-bioreactive probes, and matrix-assisted laser desorption/ionization mass spectrometry is an effective way to find and characterize phosphoproteins, known and unknown, in complex mixtures. Facile hydrolysis of human salivary phosphoproteins by the bioreactive probes was readily observed.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. B. Goshe Characterizing phosphoproteins and phosphoproteomes using mass spectrometry Brief Funct Genomic Proteomic, February 1, 2006; 4(4): 363 - 376. [Abstract] [Full Text] [PDF]