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. 2018 May 28;7(2):33. doi: 10.3390/biology7020033

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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DdRabS upregulates the secretion of lysosomal glycosidase and glucosidase enzymes. Graphical representation with standard error of the secretion of the lysosomal enzyme β-glucosidase. (A–E) Logarithmically growing cells were grown under standard secretion conditions, separated into cellular and media fractions by centrifugation, lysed with Triton X-100, and each fraction was assayed for (A) α-mannosidase (B) N-acetylglucosaminidase, (C) β-galactosidase, (D) acid phosphatase activity and (E) β-glucosidase. Symbols are as follows: (♦) AX4, (■) pDneo2a-GFP, (▲) DdRabS, and () DdRabS(DN). DdRabS overexpressing and DdRabS(DN) cells show abnormal secretion of all enzymes assayed. Secretion of the lysosomal enzyme β-glucosidase is increased in DdRabS overexpressing and DdRabS(DN) cell lines. Cells overexpressing the DdRabS protein had a higher rate of secretion of β-glucosidase compared to the WT AX4 cells. DdRabS overexpressing cells reached 100% secreted enzyme after 280 min whereas the AX4 cells peaked at 80% secretion at 280 min. The DdRabS(DN) cell line has increased secretion of β-glucosidase early on, but by 120 min, secretion is only slightly more than the WT AX4 cell line. Data represent the mean of three independent experiments. Data was run through a 2-way ANOVA to test for significance at p < 0.05.

Inline graphic — DdRabS upregulates the secretion of lysosomal glycosidase and glucosidase enzymes. Graphical representation with standard error of the secretion of the lysosomal enzyme β-glucosidase. (A–E) Logarithmically growing cells were grown under standard secretion conditions, separated into cellular and media fractions by centrifugation, lysed with Triton X-100, and each fraction was assayed for (A) α-mannosidase (B) N-acetylglucosaminidase, (C) β-galactosidase, (D) acid phosphatase activity and (E) β-glucosidase. Symbols are as follows: (♦) AX4, (■) pDneo2a-GFP, (▲) DdRabS, and () DdRabS(DN). DdRabS overexpressing and DdRabS(DN) cells show abnormal secretion of all enzymes assayed. Secretion of the lysosomal enzyme β-glucosidase is increased in DdRabS overexpressing and DdRabS(DN) cell lines. Cells overexpressing the DdRabS protein had a higher rate of secretion of β-glucosidase compared to the WT AX4 cells. DdRabS overexpressing cells reached 100% secreted enzyme after 280 min whereas the AX4 cells peaked at 80% secretion at 280 min. The DdRabS(DN) cell line has increased secretion of β-glucosidase early on, but by 120 min, secretion is only slightly more than the WT AX4 cell line. Data represent the mean of three independent experiments. Data was run through a 2-way ANOVA to test for significance at p < 0.05.