Clinical Chemistry
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Clinical Chemistry 45: 21-28, 1999;
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(Clinical Chemistry. 1999;45:21-28.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Posttranslational Deamidation of Proteins: The Case of Hemoglobin J Sardegna [{alpha}50(CD8)His->Asn->Asp]

Renata Paleari1, Elisabetta Paglietti2, Andrea Mosca1,a, Michele Mortarino3, Liliana Maccioni2, Stefania Satta2, Antonio Cao2 and Renzo Galanello2

1 Dipartimento di Scienze e Tecnologie Biomediche, Laboratorio Interdisciplinare Technologie Avanzare (L.I.T.A.), Università degli Studi, I-20090 Segrate, Milan, Italy.

2 Istituto di Clinica e Biologia dell'Età Evolutiva, Università degli Studi, I-09131 Cagliari, Italy.

3 Istituto di Fisiologia Veterinaria e Biochimica, Università degli Studi, I-20133 Milano, Italy.
a Address correspondence to this author at: L.I.T.A., Via Fratelli Cervi 93, I-20090 Segrate, Milano, Italy. Fax 39 2 2642 3302; e-mail andrea.mosca{at}unimi.it.

Hemoglobin J Sardegna [{alpha}50(CD8)His->Asn ->Asp] is a human Hb variant in which a posttranslational deamidation process takes place, transforming an Asn to an Asp residue. This variant, particularly widespread in northern Sardinia, has for the first time been characterized at the DNA level (codon 50 C->A) on the selectively amplified {alpha}2-globin gene. We determined the protein and DNA sequences and performed cellulose acetate electrophoresis, isoelectric focusing, globin chain separation, stability tests with isopropanol and heat precipitation, and oxygen affinity analyses on whole blood to fully characterize the variant. A comprehensive review of the deamidation processes involving Asn and Gln residues in mutant proteins is reported, together with a discussion of the molecular mechanisms of such deamidations. Finally, examples of other proteins of clinical importance in which Asn or Gln residues have been implicated by DNA analysis alone are presented. These findings point out the importance of the complete characterization of variant proteins by use of both DNA and protein analyses.




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