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Blood, 15 June 2008, Vol. 111, No. 12, pp. 5712-5720. Prepublished online as a Blood First Edition Paper on January 24, 2008; DOI 10.1182/blood-2007-11-122457. This Article Full Text (PDF) Supplemental Figures All Versions of this Article: blood-2007-11-122457v1 111/12/5712    most recent Alert me when this article is cited Alert me if a correction is posted 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 Gaetani, M. Articles by Speicher, D. W Search for Related Content PubMed PubMed Citation Articles by Gaetani, M. Articles by Speicher, D. W Related Collections Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted November 8, 2007 Accepted January 15, 2008 Structural and functional effects of hereditary hemolytic anemia-associated point mutations in the alpha spectrin tetramer site Massimiliano Gaetani, Sara Mootien, Sandra Harper, Patrick G Gallagher, and David W Speicher* Proteomics Laboratory, The Wistar Institute, Philadelphia, PA, United States Yale University School of Medicine, New Haven, CT, United States * Corresponding author; email: speicher{at}wistar.org. The most common hereditary elliptocytosis (HE) and hereditary pyropoikilocytosis (HPP) mutations are -spectrin missense mutations in the dimer-tetramer self-association site. In this study we systematically compared structural and functional properties of the 14 known HE/HPP mutations located in the -spectrin tetramer binding site. All mutant spectrin recombinant peptides were well folded, stable structures with only the R34W mutant exhibiting a slight structural destabilization. In contrast, binding affinities measured by isothermal titration calorimetry were greatly variable, ranging from no detectable binding observed for I24S, R28C, R28H, R28S and R45S to approximately wild type binding for R34W and K48R. Binding affinities for the other seven mutants were reduced by approximately 10 to 100-fold relative to wild type binding. Some sites, such as R28, were hot spots that were very sensitive to even relatively conservative substitutions, while other sites were only moderately perturbed by non-conservative substitutions. The R34W and K48R mutations were particularly intriguing mutations that apparently either destabilize tetramers through mechanisms not probed by the univalent tetramer binding assay or represent polymorphisms rather than the pathogenic mutations responsible for observed clinical symptoms. All 0 HE/HPP mutations studied here appear to exert their destabilizing effects through molecular recognition rather than structural mechanisms. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: Mechanisms of elliptocytosis: significant spectrin substitutions Anthony J. Baines Blood 2008 111: 5417. [Full Text] [PDF] This article has been cited by other articles: N. Mohandas and P. G. Gallagher Red cell membrane: past, present, and future Blood, November 15, 2008; 112(10): 3939 - 3948. [Abstract] [Full Text] [PDF]

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