Relationship between beta4 hydrogen bond and beta6 hydrophobic interactions during aggregate, fiber or crystal formation in oversaturated solutions of hemoglobin A and S.
Oversaturated deoxy-alpha(2)beta(2)(T4V) aggregated instantly without a delay time, which is in contrast to the delay time before the generation of fibers of deoxy-HbS and deoxy-alpha(2)beta(2)(E6V,D73H). Solubility of deoxy-alpha(2)beta(2)(T4V) was approximately 10-fold lower than that of deoxy-HbS and was similar to oxy- and deoxy-alpha(2)beta(2)(E6V,T4V). These results indicate that beta4Val in HbA in the oxy and deoxy forms with or without beta6Val facilitates hydrophobic interaction of the A-helix with the EF helix of adjacent molecules without forming a beta4/beta73 hydrogen bond. Deoxy-HbA generated crystals following aggregation as does HbC-Harlem(alpha(2)beta(2)(E6V,D73N)), while alpha(2)beta(2)(T4V) and alpha(2)beta(2)(D73H) as well as HbS, alpha(2)beta(2)(E6V,D73H) and alpha(2)beta(2)(E6V,T4V) in the oxy and deoxy forms did not form crystals, indicating in addition to the strength of beta6 amino acid hydrophobicity that the synergism between the beta4Thr hydrogen bond and beta6 hydrophobic interaction free energies on the A-helix play a critical role in formation of fibers versus crystalline nuclei during phase transition.
Adachi
Kazuhiko
K
Division of Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA. adachi@email.chop.edu
Ding
Min
M
Asakura
Toshio
T
Surrey
Saul
S
D014644:Genetic Variation; D006441:Hemoglobin A / Q000737:chemistry* / Q000235:genetics; D006451:Hemoglobin, Sickle / Q000737:chemistry* / Q000235:genetics; D006454:Hemoglobins / Q000737:chemistry; D006456:Hemoglobinuria; D006801:Humans; D006860:Hydrogen Bonding; D016296:Mutagenesis; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility; D013816:Thermodynamics
D006451:Hemoglobin, Sickle; D006454:Hemoglobins; D011994:Recombinant Proteins; C038875:deoxyhemoglobin, sickle; C100003:deoxyhemoglobin; D006441:Hemoglobin A