The cerebroside sulfate activator from pig kidney: Derivitization, cerebroside sulfate binding, and metabolic correction

Fluharty, Arvan L.; Meek, William E.; Katona, Zoltan; Tsay, Katherine K.

doi:10.1016/0885-4505(92)90010-v

Cited by 10 publications

(10 citation statements)

References 21 publications

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“…The same results were obtained with both preparations. This is in agreement with previous findings showing that Sap B isolated from pig kidney has the same activity as Sap B isolated from human tissues and is able to correct the CS hydrolytic defect in Sap B-deficient human fibroblasts (7). All later studies were thus performed with Sap B purified from pig kidney.…”

Section: Sap B-induced Solubilization Of Sls and Phospholipids From Luvssupporting

confidence: 91%

“…The Sap B molecular form in solution is almost exclusively dimeric (6,7). Crystallographic analysis has confirmed the dimer structure of Sap B (8).…”

Section: Discussionmentioning

confidence: 73%

“…In this article, we provide biochemical evidence that Sap B, a glycoprotein shown to bind and transfer SLs (7,(9)(10)(11)(12)25), binds and transfers phospholipids as well. In the past, the formation of Sap B-SL complexes has usually been analyzed using micellar dispersions of pure SLs.…”

Section: Discussionmentioning

confidence: 99%

“…Sap B, like the other three saposins, consists of z80 amino acids, including six cysteines (2), forming three disulfide bridges (5). In solution, Sap B is present as a dimer, which gives rise to higher order aggregates on increasing Sap B concentration (6,7). Crystallization has confirmed the dimer structure of Sap B, which consists of two clasped V-shaped monomers (8).…”

mentioning

confidence: 83%

“…The formation of water-soluble complexes between Sap B and SLs such as GM1 and CS has been assessed previously by a variety of methods, such as gel permeation, ultracentrifugation, and electrophoresis (7,9,12). In those experiments, the SLs were usually presented to Sap B as micelles.…”

Section: Sap B-induced Solubilization Of Sls and Phospholipids From Luvsmentioning

confidence: 99%

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Saposin B binds and transfers phospholipids

Ciaffoni

Tatti

Boe

et al. 2006

Journal of Lipid Research

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Saposin B (Sap B) is a member of a family of four small glycoproteins, Sap A, B, C, and D. Like the other three saposins, Sap B plays a physiological role in the lysosomal degradation of sphingolipids (SLs). Although the interaction of Sap B with SLs has been investigated extensively, that with the main membrane lipid components, namely phospholipids and cholesterol (Chol), is scarcely known. Using large unilamellar vesicles (LUVs) as membrane models, we have now found that Sap B simultaneously extracts from the lipid surface neutral [phosphatidylcholine (PC)] and anionic [phosphatidylinositol (PI)] phospholipids, fewer SLs [ganglioside GM1 (GM1) or cerebroside sulfate (CS)], and no Chol. More PI than SL (GM1 or CS) was solubilized from LUVs containing equal amounts of PI and SLs. An increase in PI level had a poor effect on the Sap B-induced solubilization of GM1 or CS but strongly inhibited that of PC. Sap B was able not only to bind, but also to transfer phospholipids between lipid surfaces. Both the phospholipid binding and transfer activities were optimal at low pH values. These results represent the first biochemical analysis of the Sap B interaction with phospholipids.

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Section: Sap B-induced Solubilization Of Sls and Phospholipids From Luvssupporting

confidence: 91%

“…The Sap B molecular form in solution is almost exclusively dimeric (6,7). Crystallographic analysis has confirmed the dimer structure of Sap B (8).…”

Section: Discussionmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 83%

Section: Sap B-induced Solubilization Of Sls and Phospholipids From Luvsmentioning

confidence: 99%

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Saposin B binds and transfers phospholipids

Ciaffoni

Tatti

Boe

et al. 2006

Journal of Lipid Research

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Cerebroside sulfate activator protein (Saposin B): chromatographic and electrospray mass spectrometric properties

Faull¹,

Whitelegge²,

Higginson³

et al. 1999

J. Mass Spectrom.

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Hydrogen-deuterium exchange signature of porcine cerebroside sulfate activator protein

et al. 2000

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Hydrogen–deuterium exchange can be a sensitive indicator of protein structural integrity. Comparisons were made between cerebroside sulfate activator protein (CSAct) in the native state and after treatment with guanidine hydrochloride plus dithiothreitol. Native protein has three internal disulfide bonds and treated protein has no internal disulfide bonds. The comparisons were made using hydrogen–deuterium exchange measured by electrospray ionization mass spectrometry, percentage α‐helical content measured by circular dichroism and biological activity measured by the ability to support arylsulfatase A‐catalyzed sulfate hydrolysis from cerebroside sulfate. In acidic solvent native protein has 59 exchange refractory protons and treated protein has 20 exchange refractory protons (44 and 14% of the exchangeable proton populations, respectively). In native protein the size of the exchange refractory proton population is sensitive to changes in pH, temperature and the presence of a ligand. It is uninfluenced by the presence or absence of glycosyl groups attached to Asn21. Helical content is virtually identical in native and treated protein. Biological activity is significantly reduced but not obliterated in treated protein. The hydrogen–deuterium exchange profile appears to be a sensitive signature of the correctly folded protein, and reflects a dimension of the protein structure that is not apparent in circular dichroic spectra or in the ability of the protein to support arylsulfatase A‐catalyzed sulfate hydrolysis from sulfatide. The hydrogen–deuterium exchange profile will be a valuable criterion for characterizing mutant forms of CSAct produced by recombinant and synthetic paradigms and also the native and mutant forms of related proteins. Copyright © 2000 John Wiley & Sons, Ltd.

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The cerebroside sulfate activator from pig kidney: Derivitization, cerebroside sulfate binding, and metabolic correction

Cited by 10 publications

References 21 publications

Saposin B binds and transfers phospholipids

Saposin B binds and transfers phospholipids

Cerebroside sulfate activator protein (Saposin B): chromatographic and electrospray mass spectrometric properties

Hydrogen-deuterium exchange signature of porcine cerebroside sulfate activator protein

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