Crystallization and preliminary characterization of three different crystal forms of human saposin C heterologously expressed in Pichia pastoris

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Robert Schultz-HeienbrokTimm Maier

Abstract

The amphiphilic saposin proteins (A, B, C and D) act at the lipid-water interface in lysosomes, mediating the hydrolysis of membrane building blocks by water-soluble exohydrolases. Human saposin C activates glucocerebrosidase and beta-galactosylceramidase. The protein has been expressed in Pichia pastoris, purified and crystallized in three different crystal forms, diffracting to a maximum resolution of 2.5 A. Hexagonal crystals grew from 2-propanol-containing solution and contain a single molecule in the asymmetric unit according to the Matthews coefficient. Orthorhombic and tetragonal crystals were both obtained with pentaerythritol ethoxylate and are predicted to contain two molecules in the asymmetric unit. Attempts to determine the respective crystal structures by molecular replacement using either the known NMR structure of human saposin C or a related crystal structure as search models have so far failed. The failure of the molecular-replacement method is attributed to conformational changes of the protein, which are known to be required for its biological activity. Crystal structures of human saposin C therefore might be the key to mapping out the conformational trajectory of saposin-like proteins.

References

Jun 5, 1992·Biochimica Et Biophysica Acta·W Fürst, K Sandhoff
Apr 28, 1995·The Journal of Biological Chemistry·A M VaccaroP Pucci
Jun 1, 1993·Archives of Biochemistry and Biophysics·M HiraiwaY Kishimoto
Jan 1, 1994·Human Mutation·M Horowitz, A Zimran
Oct 23, 1997·Nature Structural Biology·E LiepinshG Otting
May 18, 2001·The Journal of Biological Chemistry·X Qi, G A Grabowski
Sep 22, 2001·Acta Crystallographica. Section D, Biological Crystallography·R J Read
Dec 19, 2002·Journal of Molecular Biology·Daniel H AndersonDavid Eisenberg
Jan 9, 2003·Proceedings of the National Academy of Sciences of the United States of America·Victoria E AhnGilbert G Privé
Feb 17, 2005·Journal of Molecular Biology·Cheryl A HawkinsNico Tjandra

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Citations

Feb 25, 2010·Journal of Lipid Research·Misbaudeen Abdul-HammedKonrad Sandhoff
Oct 20, 2009·FEBS Letters·Thomas Kolter, Konrad Sandhoff
Apr 6, 2011·Protein Expression and Purification·Marialetizia MottaRosa Salvioli
Jan 1, 2007·Prion·Déborah TribouillardMarc Blondel
Nov 7, 2007·Biochemistry·Daniel StokeleyMark S P Sansom

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