Electrostatic modification at the amino termini of hemoglobin A.
The structural perturbations of the amino-terminal domains of hemoglobin A resulting from the carbamino adduct formation (a reversible in vivo electrostatic modification reaction) at Val-1(alpha) and Val-1(beta) is expected to be mimicked in the derivatives of HbA reductively alkylated at its alpha-amino terminus with aldehydes containing a negatively charged functional group at their distal end (double-headed reagents). Derivatives of HbA with galacturonic acid linked through alkylamino linkage either to Val-1(alpha) or Val-1(beta) (disubstituted derivatives) as well as to both Val-1(alpha) and Val-1(beta) (tetrasubstituted derivative) have been now prepared. All the three derivatives exhibit normal cooperativity but reduced O2 affinities. The functional consequence of the modification of HbA at its amino termini with D-galacuronic acid has been compared with that of the carboxymethylation of HbA at the same sites. This comparative study suggests that the stereochemistry of the carboxylate ion introduced into beta-cleft of Hb dictates the level of reduction in the O2 affinity of the molecule seen on derivatization. However such a unique stereochemistry of the carboxylate ion of the reagent does not appear to be crucial to lower oxygen affinity when the modification is at the amino terminus of the alpha-chain. The molecular modeling studies demonstrate that the carboxylate ion of the carbamino adduct at the amino terminus of the beta-chain as well as the carboxylate of carboxymethyl group at the same site are in a geometrical orientation that favors the formation of an intrachain ionic interaction with the epsilon-amino group of Lys-82(beta). On the other hand the stereochemistry of a carboxylate ion of galacuronic acid on Val-1(beta) appears to be appropriate to form either an intrachian salt bridge with epsilon-amino group of Ly2-82(beta) of the same chain (intrachain) or alternatively an interchain salt bridge involving the epsilon-amino group of Lys-82(beta) of the trans chain. We speculate that the latter, i.e. trans configuration is favored as a result of the potential of D-galacturonic acid bound to Val-1(beta) to form an additional hydrogen bond with trans His-143(beta).
Acharya
A S
AS
Department of Medicine, Albert Einstein College of Medicine, Bronx, New York 10461.
Bobelis
D J
DJ
White
S P
SP
D000478:Alkylation; D000595:Amino Acid Sequence; D002848:Chromatography, DEAE-Cellulose; D018360:Crystallography, X-Ray; D006441:Hemoglobin A / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D008958:Models, Molecular; D010108:Oxyhemoglobins / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D014633:Valine
D006603:Hexuronic Acids; D046911:Macromolecular Substances; D010108:Oxyhemoglobins; C007819:galacturonic acid; D006441:Hemoglobin A; C029774:oxyhemoglobin A; D014633:Valine