Biological Sciences (61) This project introduces mass spectrometry-based protein identification into an upper division biochemistry laboratory, a course required of all Biochemistry majors and an elective for Biology majors. Students are using affinity and ion exchange chromatography to isolate Bowman-Birk inhibitors, a set of stable serine protease inhibitors found in abundance in easily available seeds. Student groups are performing in-gel proteolytic digestion on the isolated inhibitors, identifying their peptide sequence by peptide mass fingerprinting and tandem mass spectrometry (MS/MS), and, once the peptide sequences are identified, accessing the entire sequence and determining the susceptible peptide bonds in each conserved domain that distinguish inhibitors of trypsin, chymotrypsin and elastase from one another. Students then are testing their hypotheses concerning essential bonds by measuring protease activity that is lost by preincubation with inhibitor. The project is reinforcing students' learning of protein structure-function and evolutionary relationships and includes production of freely available online learning materials on protein identification. The effectiveness of the course is being assessed through pre- and post- tests of students' ability to apply their newly learned concepts as compared to a control group of students who are learning the same concepts in a lecture-discussion format.
The intellectual outcomes of this project stem from its introduction of forefront techniques into an undergraduate laboratory designed to encourage student understanding of basic concepts concerning protein structure and function and of the appropriate use of modern proteomic related techniques.
The broader impacts of this project include the fact the course and the disseminated material stemming from it are designed so that they can be adopted in whole or in part by a variety of institutions. In addition, through involvement with a program that serves underrepresented minorities (the NIH Bridges to the Baccalaureate Program) the Principal Investigators are actively encouraging women and underrepresented minority students to enroll in the course.
