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J Mol Biol. 2016 Jul 31;428(15):3026-42. doi: 10.1016/j.jmb.2016.06.012. Epub 2016 Jun 24.

Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis.

Author information

1
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
2
Drug Discovery, Ontario Institute for Cancer Research, Toronto, Ontario, Canada; Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario, Canada.
3
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Molecular Structure and Function, Hospital for Sick Children, Toronto, Ontario, Canada.
4
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada; Princess Margaret Cancer Centre, Toronto, Ontario, Canada. Electronic address: prive@uhnres.utoronto.ca.

Abstract

Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction.

KEYWORDS:

Lipid hydrolase; Niemann-Pick disease; interfacial catalysis; saposin; sphingomyelin

PMID:
27349982
DOI:
10.1016/j.jmb.2016.06.012
[Indexed for MEDLINE]

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