Table 3 Thermodynamic parameters of Mg²⁺, Ca²⁺, and Zn²⁺ ion complexation with nesfatin-3s derived from ITC data

Model

Single site

Sequential two site

Single site and two independent sites

Single site

Single site, linear blank

[nesfatin-3] (µM)

120

95

120

85

80

[M²⁺] (µM)

2000

2000

2000

2000

2000

n₁

2.30 ± 0.02

–

1.01 ± 0.01

2.08 ± 0.04

0.99 ± 0.02

K_d1 (µM)

2.4 ± 0.4

116 ± 81

7 × 10^–3 ± 24 × 10^–3

9 ± 1

7 ± 1

ΔH₁ (kcal/mol)

3.47 ± 0.05

-6 ± 2

-4.57

-8.3 ± 0.2

-9.8 ± 0.4

TΔS₁ (cal/mol)

11,241

-187

6541

-1374

-2736

Intercept (µcal)

–

–

–

–

26 ± 2

Slope

–

–

–

–

-0.14 ± 0.05

n₂

–

–

1.35 ± 0.06

–

–

K_d2 (µM)

–

45.9 ± 16.1

0.42 ± 0.29

–

–

ΔH₂ (kcal/mol)

–

-6.36 ± 2.06

0.28 ± 0.87

–

–

TΔS₂ (cal/mol)

–

-437

8968.35

–

–

n₃

–

–

1.12 ± 0.19

–

–

K_d3

–

–

31.11 ± 9.17

–

–

ΔH₃ (kcal/mol)

–

–

-7.414 ± 1.83

–

–

TΔS₃ (cal/mol)

–

–

-1266

–

–