The N-terminal 18 amino acid sequence of the ?-chain of hemoglobin, as far as the end of the A helix, has been replaced by the corresponding sequence of the (-chain of fetal hemoglobin with the remaining sequence of the ?-chain retained (helices B through H). The (-?chain had the correct mass and its entire sequence was established by mass spectrometric analysis of its tryptic peptides; the ?-chain also had the correct mass. This recombinant hemoglobin (named Hb Felix) retains cooperatively and has an oxygen affinity like that of HbA both in the presence and absence of the allosteric regulators, 2,3-DPG or chloride but differs from HbF in its 2,3-DPG response. However, Hb Felix has some features that resemble fetal hemoglobin, i.e., its significantly decreased tetramer-dimer dissociation and its circular dichroism spectrum, which measure the strength of the tetramer-dimer interface in the oxy conformation and its rearrangement to the deoxy conformation, respect ively. Ev en though Hb Felix contains the HbA amino acids at its tetramer-dimer interface, which is located at a distance from the substitution sites, its interface properties resemble those of cont... HbF. Therefore, the N-terminal sequence and not just those amino acids directly involved at the subunit interface contacts with (-chains must have a strong influence on this region of the molecule. The results reinforce the concept of fluid long range relationships among various parts of the hemoglobin tetramer (Dumoulin et. al, J. Biol. Chem. 272, 31326 (1997) and demonstrate the importance of the N-terminal sequence, especially in some mutant hemoglobins, in influencing its overall structure by affecting the relationship between helices. A paper describing this work is in progress.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000862-26
Application #
6118302
Study Section
Project Start
1998-12-10
Project End
1999-11-30
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
26
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Rockefeller University
Department
Type
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
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