UF CHM 6304 - BBA 2005 Lorigan SapC 31P and 2H NMR (9 pages)

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BBA 2005 Lorigan SapC 31P and 2H NMR



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BBA 2005 Lorigan SapC 31P and 2H NMR

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Lecture Notes


Pages:
9
School:
University of Florida-Gainesville
Course:
Chm 6304 - Biomembranes
Biomembranes Documents

Unformatted text preview:

Biochimica et Biophysica Acta 1717 2005 58 66 http www elsevier com locate bba Structural changes in a binary mixed phospholipid bilayer of DOPG and DOPS upon saposin C interaction at acidic pH utilizing 31P and 2H solid state NMR spectroscopy Shadi Abu Baker a Xiaoyang Qi b Justin Newstadt a Gary A Lorigan a b a Department of Chemistry and Biochemistry Miami University Oxford OH 45056 USA Division and Program of Human Genetics Cincinnati Children s Hospital Medical Center and University of Cincinnati College of Medicine Cincinnati OH 45229 USA Received 15 July 2005 received in revised form 7 September 2005 accepted 12 September 2005 Available online 10 October 2005 Abstract Saposin C Sap C is known to stimulate the catalytic activity of the lysosomal enzyme glucosylceramidase GCase that facilitates the hydrolysis of glucosylceramide to ceramide and glucose Both Sap C and acidic phospholipids are required for full activity of GCase In order to better understand this interaction mixed bilayer samples prepared from dioleoylphosphatidylglycerol DOPG and dioleoylphosphatidylserine DOPS 5 3 ratio and Sap C were investigated using 2H and 31P solid state NMR spectroscopy at temperatures ranging from 25 to 50 C at pH 4 7 The Sap C concentrations used to carry out these experiments were 0 mol 1 mol and 3 mol with respect to the phospholipids The molecular order parameters SCD were calculated from the dePaked 2H solid state NMR spectra of Distearoyl d70 phosphatidylglycerol DSPGd70 incorporated with DOPG and DOPS binary mixed bilayers The SCD profiles indicate that the addition of Sap C to the negatively charged phospholipids is concentration dependent SCD profiles of 1 mol of the Sap C protein show only a very slight decrease in the acyl chain order However the SCD profiles of the 3 mol of Sap C protein indicate that the interaction is predominantly increasing the disorder in the first half of the acyl chain near the head group C1 C8 indicating that the amino and the



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