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Identifying a Protein by MALDI-TOF Mass Spectrometry: An Experiment for the Undergraduate Laboratory
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Abstract
Matrix-assisted laser desorption ionization time-of-flight (MALDI–TOF) mass spectrometry has become a valuable tool for performing routine biochemical analyses. A common procedure for protein identification involves using tryptic digestion to obtain masses of individual peptides derived from the protein. The masses are compared against an online database, and probability-based scoring systems are used to determine the closest protein matches. This article describes an experiment we have developed for an undergraduate honors general chemistry laboratory to introduce students to state-of-the-art mass spectrometric methods. Students are given an overview of the theory and instrumentation associated with MALDI–TOF, and gain hands-on experience with Internet tools for protein identification using mass spectral data. The experiment would be suitable for upper-division undergraduate laboratory courses as well; appropriate modifications for this purpose are also described.
Keywords (Audience):
Second-Year UndergraduateKeywords (Domain):
Analytical ChemistryKeywords (Pedagogy):
Internet / Web-Based LearningKeywords (Subject):
Mass SpectrometryCiting Articles
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This article has been cited by 7 ACS Journal articles (5 most recent appear below).
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Dale A. Rosado, Jr., Tina S. Masterson, and Douglas S. MastersonJournal of Chemical Education2011 88 (2), 178-183Mass spectrometry (MS) has been gaining in popularity in recent years owing in large part to the development of soft-ionization techniques such as matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI). These soft-ionization ...
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Danny H. Kim, Curtis D. Eckhert, Kym F. FaullJournal of Chemical Education2011 88 (1), 106-110Mass spectrometry (MS) is a powerful analytical technique that is now widely used in the chemical, physical, engineering, and life sciences, with rapidly growing applications in many areas including clinical, forensic, pharmaceutical, and environmental ...
Identification of Fatty Acids, Phospholipids, and Their Oxidation Products Using Matrix-Assisted Laser Desorption Ionization Mass Spectrometry and Electrospray Ionization Mass Spectrometry
Christopher W. Harmon, Stephen A. Mang, John Greaves and Barbara J. Finlayson-PittsJournal of Chemical Education2010 87 (2), 186-189Identification of Fatty Acids, Phospholipids, and Their Oxidation Products Using Matrix-Assisted Laser Desorption Ionization Mass Spectrometry and Electrospray Ionization Mass Spectrometry
Christopher W. Harmon, Stephen A. Mang, John Greaves and Barbara J. Finlayson-PittsJournal of Chemical Education2010 87 (2), 186-189Electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) have found increasing application in the analysis of biological samples. Using these techniques to solve problems in analytical ...
Illustrating the Concepts of Isotopes and Mass Spectrometry in Introductory Courses: A MALDI-TOF Mass Spectrometry Laboratory Experiment
Nancy Carter Dopke and Timothy Neal LovettJournal of Chemical Education2007 84 (12), 1968Illustrating the Concepts of Isotopes and Mass Spectrometry in Introductory Courses: A MALDI-TOF Mass Spectrometry Laboratory Experiment
Nancy Carter Dopke and Timothy Neal LovettJournal of Chemical Education2007 84 (12), 1968Mass spectrometry is a widely used and versatile tool for scientists in many different fields. Soft ionization techniques such as matrix-assisted laser desorption/ionization (MALDI) allow for the analysis of biomolecules, polymers, and clusters. This ...
Introduction to Biological Mass Spectrometry: Determining Identity and Species of Origin of Two Proteins
Arieh Cohen , Curt T. Reimann , Axel Mie and Carina NilssonJournal of Chemical Education2005 82 (8), 1215Introduction to Biological Mass Spectrometry: Determining Identity and Species of Origin of Two Proteins
Arieh Cohen , Curt T. Reimann , Axel Mie and Carina NilssonJournal of Chemical Education2005 82 (8), 1215We describe a laboratory exercise that is designed to give advanced undergraduate students in analytical chemistry a meaningful introduction to biological mass spectrometry. We ask the students to solve a mystery: given two proteins, what are they, and ...
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- Received: August 03, 2009
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