Porcine cerebroside sulfate activator (saposin B) secondary structure: CD, FTIR, and NMR studies

Molecular Genetics and Metabolism
A J WaringA L Fluharty

Abstract

Cerebroside sulfate activator protein (CSAct or saposin B) is one of a group of heat stable, low-molecular-weight proteins that appear to share a common structural motif. These have been referred to as saposin-like proteins and are thought to share a multiple amphipathic helical barrel structure with a conserved pattern of disulfide linkages. Porcine kidney CSAct was prepared in high purity and consisted of three major glycosylated subforms. The protein was studied by physical-chemical methods and evaluated by various methods for structural prediction. All suggest that CSAct has high amounts of alpha-helical conformation and little if any beta-sheet. Circular dichroism (CD) studies indicate 45-50% helical conformation depending on buffer and temperature. There was only a moderate loss in helical content with increasing temperature and no indication of thermal denaturation. Fourier transform infrared spectroscopy (FTIR) measurements on deuterium hydrated self-films also indicated a predominantly helical structure. Helical axis orientation was investigated by both oriented CD and FTIR dichroism, which suggested that the helical axes were roughly parallel and oriented along the axis of the surface on which the self-films had been ...Continue Reading

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Citations

Oct 25, 2000·Protein Science : a Publication of the Protein Society·J P WhiteleggeA L Fluharty
Jan 9, 2003·Proceedings of the National Academy of Sciences of the United States of America·Victoria E AhnGilbert G Privé
Oct 25, 2001·Proceedings of the National Academy of Sciences of the United States of America·J PiatigorskyZ Kozmik
May 8, 2000·The Journal of Peptide Research : Official Journal of the American Peptide Society·L M GordonA J Waring
Nov 24, 1999·Molecular Genetics and Metabolism·A L FluhartyK F Faull
Sep 22, 2011·Analytical Biochemistry·David D DixsonRobert P Doyle
Sep 26, 2000·Molecular Genetics and Metabolism·F J WaltherA J Waring
Apr 18, 2000·Journal of Mass Spectrometry : JMS·K F FaullA L Fluharty
Jan 3, 2003·Protein Expression and Purification·Victoria E AhnGilbert G Privé
Jul 24, 2002·Biochemistry·Josh W Kurutz, Ka Yee C Lee

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