Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1976 Feb;73(2):491–495. doi: 10.1073/pnas.73.2.491

Structure-function relations in hemoglobin as determined by x-ray absorption spectroscopy.

P Eisenberger, R G Shulman, G S Brown, S Ogawa
PMCID: PMC335935  PMID: 1061148

Abstract

Conclusions concerning the structure around the iron atom in oxy- and carbonmonoxyhemoglobin have been obtained by fluorescent x-ray absorption studies. The bis-imidazole heme complex was used as a model system of known structure. The ligated forms of hemoglobin, and cytochrome c at high pH, gave spectra which were very similar to the bis-imidazole complex, where the average Fe-N bond distance is known to be 1.98 A. By comparison it was possible to determine that the average Fe-N bond distances were 1.99 A in oxyhemoglobin, 1.98 A in carbonmonoxyhemoglobin, and 1.98 A in cytochrome c at pH less than 10.5, with an experimental accuracy of +/-0.02 A. An experimental comparison between oxy- and deoxyhemoglobin A showed much larger spectral changes than amongst the ligated forms. A comparison was made between the low oxygen affinity form of deoxy HbA and the high affinity form of doexy Hb Kempsey (alpha2beta992 Asp leads to Asn). All the spectral features coincided, allowing us to conclude that the average iron-ligand bond differences must be less than or equal to 0.02 A. Since the strain energy is proportional to the square of this displacement, we show that the strain energy at the iron is less than or equal to 4 X 10(-3) eV. This is negligible compared to the difference of binding energy of the high and low affinity forms, which is 0.15 eV, showing that the energies responsible for the increase of oxygen affinity are not localized at the heme.

Full text

PDF
491

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson L. Structures of deoxy and carbonmonoxy haemoglobin Kansas in the deoxy quaternary conformation. J Mol Biol. 1975 May 5;94(1):33–49. doi: 10.1016/0022-2836(75)90403-9. [DOI] [PubMed] [Google Scholar]
  2. Bunn H. F., Wohl R. C., Bradley T. B., Cooley M., Gibson Q. H. Functional properties of hemoglobin Kempsey. J Biol Chem. 1974 Dec 10;249(23):7402–7409. [PubMed] [Google Scholar]
  3. Collins D. M., Countryman R., Hoard J. L. Stereochemistry of low-spin iron porphyrins. I. Bis(imidazole)- , , , -tetraphenylporphinatoiron(3) chloride. J Am Chem Soc. 1972 Mar 22;94(6):2066–2072. doi: 10.1021/ja00761a045. [DOI] [PubMed] [Google Scholar]
  4. GIBSON Q. H. The photochemical formation of a quickly reacting form of haemoglobin. Biochem J. 1959 Feb;71(2):293–303. doi: 10.1042/bj0710293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hopfield J. J. Relation between structure, co-operativity and spectra in a model of hemoglobin action. J Mol Biol. 1973 Jun 25;77(2):207–222. doi: 10.1016/0022-2836(73)90332-x. [DOI] [PubMed] [Google Scholar]
  6. Huber R., Epp O., Formanek H. Structures of deoxy- and carbonmonoxy-erythrocruorin. J Mol Biol. 1970 Sep 14;52(2):349–354. doi: 10.1016/0022-2836(70)90035-5. [DOI] [PubMed] [Google Scholar]
  7. Kincaid B. M., Eisenberger P., Hodgson K. O., Doniach S. X-ray absorption spectroscopy using synchrotron radiation for structural investigation of organometallic molecules of biological interest. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2340–2342. doi: 10.1073/pnas.72.6.2340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. MONOD J., WYMAN J., CHANGEUX J. P. ON THE NATURE OF ALLOSTERIC TRANSITIONS: A PLAUSIBLE MODEL. J Mol Biol. 1965 May;12:88–118. doi: 10.1016/s0022-2836(65)80285-6. [DOI] [PubMed] [Google Scholar]
  9. Ogawa S., Mayer A., Shulman R. G. High resolution proton magnetic resonance study of the two quaternary states in fully ligated hemoglobin Kansas. Biochem Biophys Res Commun. 1972 Dec 18;49(6):1485–1491. doi: 10.1016/0006-291x(72)90507-4. [DOI] [PubMed] [Google Scholar]
  10. Ogawa S., Shulman R. G. High resolution nuclear magnetic resonance spectra of hemoglobin. 3. The half-ligated state and allosteric interactions. J Mol Biol. 1972 Sep 28;70(2):315–336. doi: 10.1016/0022-2836(72)90542-6. [DOI] [PubMed] [Google Scholar]
  11. Perutz M. F., Ladner J. E., Simon S. R., Ho C. Influence of globin structure on the state of the heme. I. Human deoxyhemoglobin. Biochemistry. 1974 May 7;13(10):2163–2173. doi: 10.1021/bi00707a026. [DOI] [PubMed] [Google Scholar]
  12. Perutz M. F. Stereochemistry of cooperative effects in haemoglobin. Nature. 1970 Nov 21;228(5273):726–739. doi: 10.1038/228726a0. [DOI] [PubMed] [Google Scholar]
  13. Shulman R. G., Eisenberger P., Blumberg W. E., Stombaugh N. A. Determination of the iron-sulfur distances in rubredoxin by x-ray absorption spectroscopy. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4003–4007. doi: 10.1073/pnas.72.10.4003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Shulman R. G., Hopfield J. J., Ogawa S. Allosteric interpretation of haemoglobin properties. Q Rev Biophys. 1975 Jul;8(3):325–420. doi: 10.1017/s0033583500001840. [DOI] [PubMed] [Google Scholar]
  15. Shulman R. G., Ogawa S., Wüthrich K., Yamane T., Peisach J., Blumberg W. E. The absence of "heme-heme" interactions in hemoglobin. Science. 1969 Jul 18;165(3890):251–257. doi: 10.1126/science.165.3890.251. [DOI] [PubMed] [Google Scholar]
  16. Tyuma I., Imai K., Shimizu K. Analysis of oxygen equilibrium of hemoglobin and control mechanism of organic phosphates. Biochemistry. 1973 Apr 10;12(8):1491–1498. doi: 10.1021/bi00732a004. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES