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. 2005 Jul 26;389(Pt 3):899–911. doi: 10.1042/BJ20041918

Figure 2. Far-UV CD spectra of AnxA13b in the absence and presence of PS unilamellar vesicles.

Figure 2

(A) Far-UV CD spectra of AnxA13b (0.3 mg/ml) without calcium (continuous line) and in the presence of 100 mM CaCl2 (broken line) or MgCl2 (dotted line) at 20 and 85 °C are shown. Spectra were registered in 20 mM Hepes, pH 7.4, containing 0.1 M NaCl. Inset: dependence of the Tm on CaCl2 concentration ranging from 0 (1 mM EGTA) to 100 mM (○), 100 mM MgCl2 (△) or NaCl with ionic strength equivalent to buffer containing 100 mM CaCl2 (NaCl*; □). Melting points represent the means±S.D. for at least two independent determinations. (B) Far-UV CD spectra of AnxA13b as described above, but in the presence of 50 nm PS unilamellar liposomes (800:1 lipid/protein molar ratio) without (continuous line) or with 750 μM CaCl2 (broken line) or MgCl2 (dotted line). Inset: apparent fraction of unfolded protein (Fapp) against temperature (open symbols, in the absence of PS; closed symbols, in the presence of PS vesicles; down triangles, EGTA; circles, CaCl2; up triangles, MgCl2; squares, NaCl).