Measurement of Changes in Heme Ruffling Caused by Heme-Degrading Enzyme, IsdG
Conference Year
January 2019
Abstract
Our project analyzed the interaction between IsdG, an enzyme involved in the pathogenesis of Staphylococcus aureus bacterium, and the iron-containing heme porphyrin complex in hemoglobin. This investigation was done in hopes to gain insight into the mechanism of heme degradation by IsdG, information that plays a critical role in the enzyme's potential as a drug target to combat S. aureus. Iron serves as an essential nutrient for the bacteria pathogenesis and is acquired by IsdG. Specific amino acids lining the enzyme's catalytic pocket interact with iron and are involved in ruffling the porphyrin complex in the process, including Tryptophan-67 (Trp, W). This project involved the mutation of Trp-67 into a less bulky residue, Phenylalanine (Phe, F) forming the W67F variant of IsdG. We investigated the structure and function of the W67F variant of IsdG and expected to see reduced heme ruffling and enzymatic turnover compared to that of wild-type IsdG. Spectroscopic techniques including UV-CD, UV/Vis and MCD spectroscopy were used to note differences in the secondary and electronic structures of W67F and WT IsdG.
Primary Faculty Mentor Name
Matthew D. Liptak
Graduate Student Mentors
Matthew Conger, Ariel Schuelke-Sanchez
Faculty/Staff Collaborators
Matthew Conger & Ariel Schuelke-Sanchez (Graduate Student Mentors)
Status
Undergraduate
Student College
College of Arts and Sciences
Program/Major
Biochemistry
Primary Research Category
Biological Sciences
Secondary Research Category
Health Sciences
Measurement of Changes in Heme Ruffling Caused by Heme-Degrading Enzyme, IsdG
Our project analyzed the interaction between IsdG, an enzyme involved in the pathogenesis of Staphylococcus aureus bacterium, and the iron-containing heme porphyrin complex in hemoglobin. This investigation was done in hopes to gain insight into the mechanism of heme degradation by IsdG, information that plays a critical role in the enzyme's potential as a drug target to combat S. aureus. Iron serves as an essential nutrient for the bacteria pathogenesis and is acquired by IsdG. Specific amino acids lining the enzyme's catalytic pocket interact with iron and are involved in ruffling the porphyrin complex in the process, including Tryptophan-67 (Trp, W). This project involved the mutation of Trp-67 into a less bulky residue, Phenylalanine (Phe, F) forming the W67F variant of IsdG. We investigated the structure and function of the W67F variant of IsdG and expected to see reduced heme ruffling and enzymatic turnover compared to that of wild-type IsdG. Spectroscopic techniques including UV-CD, UV/Vis and MCD spectroscopy were used to note differences in the secondary and electronic structures of W67F and WT IsdG.