Exploring the multiligand binding specificity of saposin B reveals two binding sites

Jay Tinklepaugh, Britannia M. Smith, Etta Hanlon, Chloe Zubieta, Fadi Bou-Abdallah, Robert Patrick Doyle

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Saposin B (SapB) is a human lysosomal protein, critical for the degradation of O-sulfogalactosylceramide (sulfatide). SapB binds sulfatide and presents it to the active site of the enzyme arylsulfatase A. Deficiency of SapB leads to fatal activator-deficient metachromatic leukodystrophy. Given the conformational flexibility and the large hydrophobic "pocket" produced upon (physiologically relevant) homodimerization, SapB may have broader substrate diversity than originally thought. Herein, we present evidence using fluorescence spectroscopy and computational docking studies that SapB binds a wide variety of ligands at KD values varying from micromolar to nanomolar, with entropy being the primary driving force. We further demonstrate, for the first time, that SapB has two binding sites that can sequentially (and in a preferred order) accommodate up to two ligands at once.

Original languageEnglish (US)
Pages (from-to)7141-7145
Number of pages5
JournalACS Omega
Volume2
Issue number10
DOIs
StatePublished - 2017

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Saposins
Binding sites
Binding Sites
Ligands
Fluorescence spectroscopy
Entropy
Enzymes
Proteins
Degradation
Substrates
Cerebroside-Sulfatase

ASJC Scopus subject areas

  • Chemical Engineering(all)
  • Chemistry(all)

Cite this

Tinklepaugh, J., Smith, B. M., Hanlon, E., Zubieta, C., Bou-Abdallah, F., & Doyle, R. P. (2017). Exploring the multiligand binding specificity of saposin B reveals two binding sites. ACS Omega, 2(10), 7141-7145. https://doi.org/10.1021/acsomega.7b01334

Exploring the multiligand binding specificity of saposin B reveals two binding sites. / Tinklepaugh, Jay; Smith, Britannia M.; Hanlon, Etta; Zubieta, Chloe; Bou-Abdallah, Fadi; Doyle, Robert Patrick.

In: ACS Omega, Vol. 2, No. 10, 2017, p. 7141-7145.

Research output: Contribution to journalArticle

Tinklepaugh, J, Smith, BM, Hanlon, E, Zubieta, C, Bou-Abdallah, F & Doyle, RP 2017, 'Exploring the multiligand binding specificity of saposin B reveals two binding sites', ACS Omega, vol. 2, no. 10, pp. 7141-7145. https://doi.org/10.1021/acsomega.7b01334
Tinklepaugh, Jay ; Smith, Britannia M. ; Hanlon, Etta ; Zubieta, Chloe ; Bou-Abdallah, Fadi ; Doyle, Robert Patrick. / Exploring the multiligand binding specificity of saposin B reveals two binding sites. In: ACS Omega. 2017 ; Vol. 2, No. 10. pp. 7141-7145.
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