Modification of human hemoglobin with methyl acyl phosphates derived from dicarboxylic acids. Systematic relationships between cross-linked structure and oxygen-binding properties.
Human hemoglobin was reacted with five dicarboxylic acid bis(methyl phosphate) reagents under different ligand conditions. The bis(methyl phosphate) reagents tested were derived from fumaric, isophthalic, terephthalic, trans-stilbene-3,3'-dicarboxylic, and trans-stilbene-4,4'-dicarboxylic acids. These acyl phosphate mixed anhydrides are anionic electrophiles and will react with N-terminal amino and lysyl epsilon-amino groups to form amides. The major and many of the minor reaction products that result have been isolated and structurally characterized by globin chain and peptide analysis. Products which are not cross-linked, intrachain linked, and interchain singly and doubly cross-linked occur in proportions which depend upon the reaction conditions and reagent. Modifications of the beta chains were limited to the amino groups of beta 1Val, beta 82Lys, and, to a minor extent, beta 144Lys. In the case of the smaller reagents, the amino groups of alpha 1Val, alpha 99Lys, and, to a minor extent, alpha 139Lys were modified. The oxygen binding affinities of most of the major modified hemoglobins have been measured and are characterized by P50 values from about 1/2 to over 5 times that of unmodified human hemoglobin. Most show strong cooperativity with Hill coefficients (n) of 2.0 or greater. Several of the products that are cross-linked between the beta 1Val of one chain and the beta 82Lys of the other chain have oxygen affinities in a physiologically useful range for oxygen transport and delivery. An inverse linear correlation has been found between the log of P50 and bridging distances for the hemoglobins cross-linked between beta 1Val of one chain and the beta 82Lys of the other chain.(ABSTRACT TRUNCATED AT 250 WORDS)
Jones
R T
RT
Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health Sciences University, Portland 97201.
Head
C G
CG
Fujita
T S
TS
Shih
D T
DT
Wodzinska
J
J
Kluger
R
R
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000737:chemistry*; D003998:Dicarboxylic Acids / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005650:Fumarates / Q000737:chemistry; D005914:Globins / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism*; D010710:Phosphates / Q000737:chemistry*; D010795:Phthalic Acids / Q000737:chemistry
D003432:Cross-Linking Reagents; D003998:Dicarboxylic Acids; D005650:Fumarates; D006454:Hemoglobins; D010710:Phosphates; D010795:Phthalic Acids; C059768:isophthalate; C011363:terephthalic acid; C032005:fumaric acid; D005914:Globins; D010100:Oxygen