A single amino acid substitution (Asp leads to Asn) in a phosphoglycerate kinase variant (PGK München) associated with enzyme deficiency.

H Fujii,W.K Krietsch,A Yoshida

doi:10.1016/s0021-9258(18)43755-6

Abstract

The structural abnormality of the phosphoglycerate kinase variant, PGK München, associated with red cell enzyme deficiency and heat instability, was elucidated by a microscale peptide-mapping method. A single amino acid substitution, from aspartic acid in the normal enzyme to asparagine in the variant enzyme, was found. From the known amino acid sequence of normal human phosphoglycerate kinase, the substitution is in the aspartic acid residue located at 268th position from the NH2-terminal of the protein.

Highlights

A Single Amino Acid Substitution(Asp + Asn) in a Phosphoglycerate
Fromthe known amino acid sequence of normal human phosphoglycerate kinase, the substitution is in the aspartic acid residue located at 268th position from
The variant phosphoglycerate kinase was purified by the affinity chromatography with ATP-ribosyl-adipoyldihydrazo-Sepharos4eB and gel filtration with Sephadex G-75 as previously reported [14, 18].The normal phosphoglycerate kinase was purified by the same method

Summary

THEJOURNAL OF BIOLOGICACHLEMISTRY

Kinase Variant (PGK Munchen) Associated with Enzyme Deficiency*. The structural abnormality of the phosphoglycerate kinase variant, PGK Munchen, associated with red cell enzyme deficiency and heat instability, was elucidated bya microscale peptide-mapping method. The peptide spots were visualizedwith 0.03%ninhydrin in acetone In contrast to these variants, PGK Munchen is not associ- 2 M acetic acid using a small funnel. The have been reported[15,16].A specific amino acid subst i tution peptides, i.e. H,N-Asp-Leu-Met-Ser-Lys-COOH and H?N-Asn-LeuMet-Ser-Lys-COOH, were synthesized by the modified Merrifield of PGK-2, which is fairly common in South Pacific popula- procedure [23].and purified by preparative electrophoresis on Whattions, has been determined [17]. RESULTSANDDISCUSSION paper reportsthe determination of the amino acid substitution The results of purification of the normal and variant enof PGK Munchen using about500 ml of the subject’s blood.

Structural Abnormality of a Phosphoglycerate KinVaasreiant

TABLEI indicating that the negative charge of the asparatyl residue

Aspartic acid