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Effects of methyl acetyl phosphate, a covalent antisickling agent, on the density profiles of sickle erythrocytes.
J Lab Clin Med. 1992 Jul; 120(1):152-8.JL

Abstract

Methyl acetyl phosphate specifically acetylates valine-1, lysine-82, and lysine-144 in the 2,3-diphosphoglycerate binding cleft of hemoglobin S, thereby inhibiting its gelation (greater than 32 gm/dl) at pH 7.4. To extend these findings, the effect of methyl acetyl phosphate on the density of sickle cells has been evaluated by phthalate ester gradient centrifugation and by Larex-Percoll density centrifugation. After treatment with methyl acetyl phosphate (40% modification of the intracellular hemoglobin S), oxygenated sickle erythrocytes had a lowered density profile, as measured in a phthalate ester gradient. Thus 83% of untreated oxygenated sickle cells had densities greater than 1.098 gm/ml, whereas after treatment with methyl acetyl phosphate, 52% of the cells were in this density range. Under anaerobic conditions, methyl acetyl phosphate was even more effective in lowering cell density. For example, 50% of untreated deoxygenated cells had densities greater than 1.098 gm/ml, but none of the cells treated with methyl acetyl phosphate were this dense. For studies with Larex-Percoll density gradients, sickle erythrocytes were first separated into two fractions (densities greater than and less than 1.1 gm/ml) by Percoll-Hypaque centrifugation. The amount of oxygenated sickle cells exhibiting densities greater than 1.074 gm/ml decreased by about 32% on treatment with methyl acetyl phosphate. For deoxygenated sickle cells, treatment with methyl acetyl phosphate resulted in an average decrease of approximately 24% in the number of cells with densities greater than 1.074 gm/ml.(

ABSTRACT

TRUNCATED AT 250 WORDS)

Authors+Show Affiliations

Rockefeller University, New York, NY 10021.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

1613320

Citation

Ueno, H, et al. "Effects of Methyl Acetyl Phosphate, a Covalent Antisickling Agent, On the Density Profiles of Sickle Erythrocytes." The Journal of Laboratory and Clinical Medicine, vol. 120, no. 1, 1992, pp. 152-8.
Ueno H, Yatco E, Benjamin LJ, et al. Effects of methyl acetyl phosphate, a covalent antisickling agent, on the density profiles of sickle erythrocytes. J Lab Clin Med. 1992;120(1):152-8.
Ueno, H., Yatco, E., Benjamin, L. J., & Manning, J. M. (1992). Effects of methyl acetyl phosphate, a covalent antisickling agent, on the density profiles of sickle erythrocytes. The Journal of Laboratory and Clinical Medicine, 120(1), 152-8.
Ueno H, et al. Effects of Methyl Acetyl Phosphate, a Covalent Antisickling Agent, On the Density Profiles of Sickle Erythrocytes. J Lab Clin Med. 1992;120(1):152-8. PubMed PMID: 1613320.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Effects of methyl acetyl phosphate, a covalent antisickling agent, on the density profiles of sickle erythrocytes. AU - Ueno,H, AU - Yatco,E, AU - Benjamin,L J, AU - Manning,J M, PY - 1992/7/1/pubmed PY - 1992/7/1/medline PY - 1992/7/1/entrez SP - 152 EP - 8 JF - The Journal of laboratory and clinical medicine JO - J Lab Clin Med VL - 120 IS - 1 N2 - Methyl acetyl phosphate specifically acetylates valine-1, lysine-82, and lysine-144 in the 2,3-diphosphoglycerate binding cleft of hemoglobin S, thereby inhibiting its gelation (greater than 32 gm/dl) at pH 7.4. To extend these findings, the effect of methyl acetyl phosphate on the density of sickle cells has been evaluated by phthalate ester gradient centrifugation and by Larex-Percoll density centrifugation. After treatment with methyl acetyl phosphate (40% modification of the intracellular hemoglobin S), oxygenated sickle erythrocytes had a lowered density profile, as measured in a phthalate ester gradient. Thus 83% of untreated oxygenated sickle cells had densities greater than 1.098 gm/ml, whereas after treatment with methyl acetyl phosphate, 52% of the cells were in this density range. Under anaerobic conditions, methyl acetyl phosphate was even more effective in lowering cell density. For example, 50% of untreated deoxygenated cells had densities greater than 1.098 gm/ml, but none of the cells treated with methyl acetyl phosphate were this dense. For studies with Larex-Percoll density gradients, sickle erythrocytes were first separated into two fractions (densities greater than and less than 1.1 gm/ml) by Percoll-Hypaque centrifugation. The amount of oxygenated sickle cells exhibiting densities greater than 1.074 gm/ml decreased by about 32% on treatment with methyl acetyl phosphate. For deoxygenated sickle cells, treatment with methyl acetyl phosphate resulted in an average decrease of approximately 24% in the number of cells with densities greater than 1.074 gm/ml.(ABSTRACT TRUNCATED AT 250 WORDS) SN - 0022-2143 UR - https://www.unboundmedicine.com/medline/citation/1613320/Effects_of_methyl_acetyl_phosphate_a_covalent_antisickling_agent_on_the_density_profiles_of_sickle_erythrocytes_ DB - PRIME DP - Unbound Medicine ER -