Uwo Thesis Binding

Uwo Thesis Binding-75
In particular, spectroscopic analysis of the YFYF mutant mixed with Hb provided evidence of a trapped heme transfer intermediate similar to that observed in the crystal structure of the wild-type protein. aureus strain RN6390 Δisd H::spec R were constructed by members of his lab. aureus USA300 strain and performed the growth experiments. 2 1.2: Human iron metabolism ........................................................................................................ 3 1.2.1: Iron storage, transport and regulation ............................................................................ .................................................................................... 5 1.2.3: Haptoglobin and hemopexin .......................................................................................... 6 1.3: Staphylococcus aureus iron acquisition systems .................................................................. In the second part of my study, Hp was found to inhibit heme transfer from Hb to Isd B in a concentration-dependent manner and heme transfer was completely blocked when Hp was added to Hb above the binding stoichiometry determined by structural and solution studies. The Isd BN1N2 (F366A) variant was cloned with assistance from William Pardoe. 9 1.3.1: Siderophore mediated iron uptake systems .................................................................... 20 2.2.1: Cloning of in vivo construct of Isd B for complementation study ................................

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Heme is then transferred to Isd A or Isd C, which then relay it to other Isd proteins for internalization. (1985) Uptake of iron from hemoglobin and the haptoglobin-hemoglobin complex by hemolytic bacteria.

Isd B contains two NEAT (NEAr Iron Transporter) domains, and both domains and the intervening linker are needed for efficient heme uptake from Hb.

To date, the Iron Surface Determinant (Isd) system is the only heme utilization pathway identified in S. Isd B is the primary receptor at the cell surface that binds Hb to extract heme. Haptoglobin blocks formation of a key intermediate in heme transfer from hemoglobin to Isd B of Staphylococcus aureus. Angelé Arrieta cloned the YFYF variant expression plasmid.

Heme is then transferred to Isd A or Isd C, which then relay it to other Isd proteins for internalization. I produced and purified YFYF protein and carried out the activity assays in collaboration with Catherine. aureus growth experiments on Hb as a sole iron source and demonstrating inhibition by Hp. iv This project required Ethics Approval for purifying hemoglobin from fresh human blood.

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Uwo Thesis Binding Emerson Essay History Summary

We use cookies to offer you a better experience, personalize content, tailor advertising, provide social media features, and better understand the use of our services.We use cookies to make interactions with our website easy and meaningful, to better understand the use of our services, and to tailor advertising.For further information, including about cookie settings, please read our Cookie Policy .To learn more or modify/prevent the use of cookies, see our Cookie Policy and Privacy Policy.Your thesis will be available online at [email protected] and on the web up to two weeks following your final submission.You retain copyright to your thesis and may make it available on a personal website and pursue other sources of publication as well.Your graduate unit may require one or more bound paper copies of your thesis.Sc., The University of Waterloo, 2014 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Microbiology and Immunology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) July 2016 © Jun Wen Li, 2016 ii ABSTRACT Staphylococcus aureus is a bacterial pathogen of major health concern. (1956) Interrelations between the catalytic function of heme proteins and the structural modifications of their protein parts. Hb released from lysed erythrocytes is quickly captured by serum haptoglobin (Hp) and the complex is degraded in the liver where released iron is then recycled. Catherine Bowden produced recombinant Isd BN1N2, performed stopped-flow experiments, crystallized the Isd BN1N2-Hb complex, and wrote the first draft of the manuscript. Anson Chan helped with solving the crystal structure. (1969) Formation of hemichromes from oxidized hemoglobin subunits. In the first part of my study, the molecular mechanism of heme transfer from Hb to Isd B was investigated using site-directed mutagenesis. Residues in the heme-binding pocket were identified by inspection of a crystal structure of the complex of Isd B and Hb.

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