This proposal involves the elucidation of structure-function relationships in recombinant transferrins, e.g., human transferrin, hen ovo-transferrin and half-molecules thereof, based on a series of site directed mutants. A major aim will be to establish by a series of physico-chemical approaches, such as NMR, fluorescence resonance energy transfer, and EPR of spin-labeled proteins, reliable indices of the """"""""open"""""""" and """"""""closed"""""""" conformations. These measurements will be undertaken as a function of pH, ionic strength, and degree of metal ligation. Metal binding affinities will be correlated with the conformational states of the proteins and various mutants. The principal criterion for metal binding affinity will be kinetic constants for the release of the bound metal ion to a competing high affinity chelator. X-ray crystallographic studies of various mutants will be continued in order to determine any marked effects in the structure of the protein arising from the mutants. Iron sequestration in blood plasma by transferrin and delivery of this iron via receptor mediated endocytosis to various tissues are biologically essential functions. This natural physiological process is also utilized in the supply of essential iron to neoplastic tissues. The fundamentals of structure-function relationships in transferrin are essential to understanding the molecular basis of certain defects in iron delivery and to the design of rational interventions in the delivery of iron to neoplasms.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK021739-16A1
Application #
2614176
Study Section
Metallobiochemistry Study Section (BMT)
Program Officer
Badman, David G
Project Start
1978-12-01
Project End
2002-06-30
Budget Start
1998-07-01
Budget End
1999-06-30
Support Year
16
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Vermont & St Agric College
Department
Biochemistry
Type
Schools of Medicine
DUNS #
066811191
City
Burlington
State
VT
Country
United States
Zip Code
05405
Yoon, Dennis J; Chen, Kevin Y; Lopes, André M et al. (2017) Mathematical modeling of mutant transferrin-CRM107 molecular conjugates for cancer therapy. J Theor Biol 416:88-98
Das, Anupam; Nag, Sagarika; Mason, Anne B et al. (2016) Endosome-mitochondria interactions are modulated by iron release from transferrin. J Cell Biol 214:831-45
Mathies, Guinevere; Gast, Peter; Chasteen, N Dennis et al. (2015) Exploring the Fe(III) binding sites of human serum transferrin with EPR at 275 GHz. J Biol Inorg Chem 20:487-96
Luck, Ashley N; Mason, Anne B (2013) Structure and dynamics of drug carriers and their interaction with cellular receptors: focus on serum transferrin. Adv Drug Deliv Rev 65:1012-9
Deblonde, Gauthier J-P; Sturzbecher-Hoehne, Manuel; Mason, Anne B et al. (2013) Receptor recognition of transferrin bound to lanthanides and actinides: a discriminating step in cellular acquisition of f-block metals. Metallomics 5:619-26
Sturzbecher-Hoehne, Manuel; Goujon, Christophe; Deblonde, Gauthier J-P et al. (2013) Sensitizing curium luminescence through an antenna protein to investigate biological actinide transport mechanisms. J Am Chem Soc 135:2676-83
Luck, Ashley N; Bobst, Cedric E; Kaltashov, Igor A et al. (2013) Human serum transferrin: is there a link among autism, high oxalate levels, and iron deficiency anemia? Biochemistry 52:8333-41
Steere, Ashley N; Chasteen, N Dennis; Miller, Brendan F et al. (2012) Structure-based mutagenesis reveals critical residues in the transferrin receptor participating in the mechanism of pH-induced release of iron from human serum transferrin. Biochemistry 51:2113-21
Steere, Ashley N; Byrne, Shaina L; Chasteen, N Dennis et al. (2012) Kinetics of iron release from transferrin bound to the transferrin receptor at endosomal pH. Biochim Biophys Acta 1820:326-33
Luck, Ashley N; Mason, Anne B (2012) Transferrin-mediated cellular iron delivery. Curr Top Membr 69:3-35

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