This research program is centered around the application of protein crystallography (a form of microscopy which uses X-rays in place of light rays) to directly produce high resolution images of protein structure. Much of our research effort has been aimed at achieving a more detailed understanding of the relationships between hemoglobin structure and function. In particular, we have determined the structures of a number of variant and chemically modified hemoglobins. By identifying the differences between the structures of these abnormal hemoglobins and normal human deoxyhemoglobin we have been able to 1) pinpoint structural features which are essential for full expression of hemoglobin function, 2) suggest a structural basis for the action of a number of drugs which have been shown to be effective in reducing the polymerizaion of sickle cell hemoglobin, and 3) uncover anion binding sites which may be important in the interaction of hemoglobin with CO2. In collaboration with Dr. David Metzler at Iowa State University we are also studying the crystal of aspartate aminotransferase purified from pig heart. Currently, we are analyzing a 2.7 A electron density map of the enzyme as well as studying complexes of the enzyme with substrates and inhibitors. Our studies should reveal structural details which will be useful in understanding the mechanism of action of this enzyme as well as of the large class of enzymes which used pyridoxal phosphate as a coenzyme. More recently, we have started studies aimed at crystallizing specific fragments of DNA and DNA-protein complexes. The work is still in its initial stages, but we hope that eventually it will add to our understanding of the structural aspects of DNA-protein interactions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
9R01GM040852-14
Application #
3298738
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1978-06-01
Project End
1993-02-28
Budget Start
1988-03-01
Budget End
1989-02-28
Support Year
14
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Iowa
Department
Type
Schools of Medicine
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242
Borgstahl, G E; Rogers, P H; Arnone, A (1994) The 1.9 A structure of deoxy beta 4 hemoglobin. Analysis of the partitioning of quaternary-associated and ligand-induced changes in tertiary structure. J Mol Biol 236:831-43
Ramaswamy, S; Kratzer, D A; Hershey, A D et al. (1994) Crystallization and preliminary crystallographic studies of Saccharomyces cerevisiae alcohol dehydrogenase I. J Mol Biol 235:777-9
Borgstahl, G E; Rogers, P H; Arnone, A (1994) The 1.8 A structure of carbonmonoxy-beta 4 hemoglobin. Analysis of a homotetramer with the R quaternary structure of liganded alpha 2 beta 2 hemoglobin. J Mol Biol 236:817-30
Kavanaugh, J S; Moo-Penn, W F; Arnone, A (1993) Accommodation of insertions in helices: the mutation in hemoglobin Catonsville (Pro 37 alpha-Glu-Thr 38 alpha) generates a 3(10)-->alpha bulge. Biochemistry 32:2509-13
Pan, Q W; Tanase, S; Fukumoto, Y et al. (1993) Functional roles of valine 37 and glycine 38 in the mobile loop of porcine cytosolic aspartate aminotransferase. J Biol Chem 268:24758-65
Kavanaugh, J S; Rogers, P H; Arnone, A (1992) High-resolution X-ray study of deoxy recombinant human hemoglobins synthesized from beta-globins having mutated amino termini. Biochemistry 31:8640-7
Vasseur, C; Blouquit, Y; Kister, J et al. (1992) Hemoglobin Thionville. An alpha-chain variant with a substitution of a glutamate for valine at NA-1 and having an acetylated methionine NH2 terminus. J Biol Chem 267:12682-91
Kavanaugh, J S; Rogers, P H; Case, D A et al. (1992) High-resolution X-ray study of deoxyhemoglobin Rothschild 37 beta Trp----Arg: a mutation that creates an intersubunit chloride-binding site. Biochemistry 31:4111-21
Silva, M M; Rogers, P H; Arnone, A (1992) A third quaternary structure of human hemoglobin A at 1.7-A resolution. J Biol Chem 267:17248-56
Taylor, J E; Metzler, D E; Arnone, A (1990) Modeling inhibitors in the active site of aspartate aminotransferase. Ann N Y Acad Sci 585:58-67

Showing the most recent 10 out of 11 publications