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. 2023 May 2;8:76. Originally published 2023 Feb 14. [Version 2] doi: 10.12688/wellcomeopenres.18937.2

Figure 4. Hyaluronidase activity of soluble Transmembrane protein 2 (sTMEM2).

Figure 4.

( A) Top, size exclusion chromatography of 100 μg/ml fluorescein-labelled high-molecular-weight Hyaluronic acid (HMW-HA) or HA 22-mer (dp22, ~5 kDa). Bottom; size exclusion chromatography of 100 μg/ml fluorescein-labelled HMW-HA incubated overnight with 100 μg/ml sTMEM2 or sperm hyaluronidase (pH 6.0, 1 mM CaCl 2). Only the sperm hyaluronidase was able to degrade HMW-HA. Shown is a representative of n = 3 experiments. ( B) Ultrafiltration of 0.1 μg/ml fluorescein-labelled HMW-HA incubated overnight with 100 μg/ml sTMEM2 and sperm hylauronidase (pH 6.0, 1 mM CaCl 2). The fluorescence was measured in the solution before and after filtration (I, input; F, filtrate). Only the sperm hyaluronidase was able to degrade HMW-HA. Data are presented as mean values ± s.e.m. for n = 3 independent experiments. ( C) Circular dichroism spectra of sTMEM2 incubated overnight at 4°C (teal) and 37°C (magenta). The minimum at 217 nm is characteristic of β-sheet structure. There is no evidence of denaturation at the higher temperature. The raw measurements are available in Underlying data.