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Blood, 15 February 2008, Vol. 111, No. 4, pp. 1972-1979. Prepublished online as a Blood First Edition Paper on December 3, 2007; DOI 10.1182/blood-2007-06-097022. This Article Full Text (PDF) All Versions of this Article: blood-2007-06-097022v1 111/4/1972    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 Kawasaki, N. Articles by Yamamoto, K. Search for Related Content PubMed PubMed Citation Articles by Kawasaki, N. Articles by Yamamoto, K. Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted June 21, 2007 Accepted November 6, 2007 The sugar-binding ability of ERGIC-53 is enhanced by its interaction with MCFD2 Norihito Kawasaki, Yoko Ichikawa, Ichiro Matsuo, Kiichiro Totani, Naoki Matsumoto, Yukishige Ito, and Kazuo Yamamoto* Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba, Japan Synthetic Cellular Chemistry Laboratory, RIKEN, The Institute of Physical and Chemical Research, Wako, Saitama, Japan CREST, JST, Wako, Saitama, Japan * Corresponding author; email: yamamoto{at}k.u-tokyo.ac.jp. Combined deficiency of factors V and VIII (F5F8D) is a bleeding disorder caused by mutations in LMAN1 or MCFD2. LMAN1 encodes ERGIC-53, a cargo receptor with an L-type lectin domain and MCFD2 is a EF-hand-containing protein. We prepared a biotinylated, soluble form of ERGIC-53, which we labeled with R-phycoerythrin conjugated streptavidin. By flow cytometry, sERGIC-53-SA bound to HeLaS3 cells in the presence of calcium but only after preincubation with MCFD2. Treating the cells with endo H or incubating them with high mannose-type oligosaccharides, especially M8B, abrogated sERGIC-53-SA binding. Surface plasmon resonance experiments demonstrated that MCFD2 specifically bound to sERGIC-53 and two MCFD2 mutants found in F5F8D patients had a Ka that was three or four orders of magnitude lower for sERGIC-53 than for wild-type MCFD2. The Ka of sERGIC-53 and MCFD2 was measured at several pH values and calcium concentrations, and we found that at a calcium concentration less than 0.2 mM, this interaction became significantly weaker. These results demonstrate that the binding of ERGIC-53 to sugar is enhanced by its interaction with MCFD2 and defects in this interaction in F5F8D patients may be the cause for reduced secretion of factors V and VIII. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: D. Hu, Y. Kamiya, K. Totani, D. Kamiya, N. Kawasaki, D. Yamaguchi, I. Matsuo, N. Matsumoto, Y. Ito, K. Kato, et al. Sugar-binding activity of the MRH domain in the ER {alpha}-glucosidase II {beta} subunit is important for efficient glucose trimming Glycobiology, October 1, 2009; 19(10): 1127 - 1135. [Abstract] [Full Text] [PDF] C. A. Bue and C. Barlowe Molecular Dissection of Erv26p Identifies Separable Cargo Binding and Coat Protein Sorting Activities J. Biol. Chem., September 4, 2009; 284(36): 24049 - 24060. [Abstract] [Full Text] [PDF]

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