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This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Maruno, M. Articles by Sassa, S. Search for Related Content PubMed PubMed Citation Articles by Maruno, M. Articles by Sassa, S. Related Collections Red Cells Clinical Trials and Observations Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 15 May 2001, Vol. 97, No. 10, pp. 2972-2978 CLINICAL OBSERVATIONS, INTERVENTIONS, AND THERAPEUTIC TRIALS Highly heterogeneous nature of -aminolevulinate dehydratase (ALAD) deficiencies in ALAD porphyria Motoyoshi Maruno, Kazumichi Furuyama, Reiko Akagi, Yutaka Horie, Kuniaki Meguro, Luba Garbaczewski, Nicholas Chiorazzi, Manfred O. Doss, Alexandre Hassoun, Rudolf Mercelis, Luc Verstraeten, Pauline Harper, Ylva Floderus, Stig Thunell, and Shigeru Sassa From Rockefeller University, New York, NY; Okayama Prefectural University, Okayama, Japan; Cornell University-Northshore Hospital, Long Island, NY; Phillips University, Marburg, Germany; Universite Catholique de Louvain, Bruxelles, Belgium; University Hospital of Antwerp, Belgium; and Huddinge University Hospital, Huddinge, Sweden. The properties of 9 -aminolevulinate dehydratase (ALAD) mutants from patients with ALAD porphyria (ADP) were examined by bacterial expression of their complementary DNAs and by enzymologic and immunologic assays. ALADs were expressed as glutathione-S-transferase (GST) fusion proteins in Escherichia coli and purified by glutathione-affinity column chromatography. The GST-ALAD fusion proteins were recognized by anti-ALAD antibodies and were enzymatically active as ALAD. The enzymatic activities of 3 ALAD mutants, K59N, A274T, and V153M, were 69.9%, 19.3%, and 41.0% of that of the wild-type ALAD, respectively, whereas 6 mutants, G133R, K59N/G133R, F12L, R240W, V275M, and delTC, showed little activity (< 8%). These variations generally reflect the phenotype of ALAD in vivo in patients with ADP and indicate that GST-ALAD fusion protein is indeed useful for predicting of the phenotype of ALAD mutants. The location of F12L mutation in the enzyme's molecular structure indicates that its disturbance of the quaternary contact of the ALAD dimer appears to have a significant influence on the enzymatic activity. Mouse monoclonal antibodies to human ALAD were developed that specifically recognized a carboxy terminal portion of ALAD, or other regions in the enzyme. This study represents the first complete analysis of 9 mutants of ALAD identified in ADP and indicates the highly heterogeneous nature of mutations in this disorder. © 2001 by The American Society of Hematology.   CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: L. Tang, S. Breinig, L. Stith, A. Mischel, J. Tannir, B. Kokona, R. Fairman, and E. K. Jaffe Single Amino Acid Mutations Alter the Distribution of Human Porphobilinogen Synthase Quaternary Structure Isoforms (Morpheeins) J. Biol. Chem., March 10, 2006; 281(10): 6682 - 6690. [Abstract] [Full Text] [PDF]

	Copyright © 2001 by American Society of Hematology         Online ISSN: 1528-0020