Abstract

The applicants long-term objective is to obtain a detailed understanding of the functioning of human embryonic haemoglobins and, in so doing, to gain an insight into the normal behavior of the oxygen transport system which operates at the earliest stages of human development. This data will provide valuable parameters related to the physiology and development of the early human individual. There are three specific aims which constitute this project namely (i) the preparation, from nonembryonic sources, of a yeast system capable of expressing, in high yields, functional forms of each of the three human embryonic haemoglobins; (ii) the isolation of purified haemoglobins to be used in oxygen binding equilibrium and kinetic investigations of the protein function; directed towards the identification of the factors controlling their interactions with and (iii) the crystallization and subsequent three dimensional structure determination of each haemoglobin so allowing the rationalization of the functional properties of these proteins at the molecular level.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK047499-02
Application #
2147159
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Project Start
1993-08-10
Project End
1996-07-31
Budget Start
1994-08-01
Budget End
1995-07-31
Support Year
2
Fiscal Year
1994
Total Cost
Indirect Cost

  Institution

Name
University of Auckland
Department
Type
DUNS #
City
Auckland
State
Country
New Zealand
Zip Code
1010

  Publications

Kidd, R D; Baker, H M; Mathews, A J et al. (2001) Oligomerization and ligand binding in a homotetrameric hemoglobin: two high-resolution crystal structures of hemoglobin Bart's (gamma(4)), a marker for alpha-thalassemia. Protein Sci 10:1739-49
Kidd, R D; Mathews, A; Baker, H M et al. (2001) Subunit dissociation and reassociation leads to preferential crystallization of haemoglobin Bart's (gamma4) from solutions of human embryonic haemoglobin Portland (zeta2gamma2) at low pH. Acta Crystallogr D Biol Crystallogr 57:921-4
Mathews, A J; Brittain, T (2001) Haem disorder in recombinant- and reticulocyte-derived haemoglobins: evidence for stereoselective haem insertion in eukaryotes. Biochem J 357:305-11
Zheng, T; Brittain, T; Watmough, N J et al. (1999) The role of amino acid alpha38 in the control of oxygen binding to human adult and embryonic haemoglobin Portland. Biochem J 343 Pt 3:681-5
Zheng, T; Zhu, Q; Brittain, T (1999) Origin of the suppression of chloride ion sensitivity in human embryonic hemoglobin Gower II. IUBMB Life 48:435-7
Sutherland-Smith, A J; Baker, H M; Hofmann, O M et al. (1998) Crystal structure of a human embryonic haemoglobin: the carbonmonoxy form of gower II (alpha2 epsilon2) haemoglobin at 2.9 A resolution. J Mol Biol 280:475-84
McLennan, A E; Brittain, T (1998) Non-synergistic interactions between strong allosteric effectors and human embryonic and adult haemoglobins. Biochem Mol Biol Int 44:175-83
Hofmann, O M; Brittain, T (1998) Partitioning of oxygen and carbon monoxide in the three human embryonic hemoglobins. Hemoglobin 22:313-9
Brittain, T; Hofmann, O M; Watmough, N J et al. (1997) A two-state analysis of co-operative oxygen binding in the three human embryonic haemoglobins. Biochem J 326 ( Pt 2):299-303
Hofmann, O M; Brittain, T; Wells, R M (1997) The control of oxygen affinity in the three human embryonic haemoglobins by respiration linked metabolites. Biochem Mol Biol Int 42:553-66

Showing the most recent 10 out of 17 publications