TABLE 5.

Identification of the binding of dockerin polypeptides to CohScaC cohesin as evaluated by native gel electrophoresis and isothermal titration calorimetry corroborated with the association and dissociation constants for the binding of DocScaB and mutant derivatives to immobilized CohScaC evaluated by surface plasmon resonance using a BIAcore system

K_D was determined as k_off/k_on. The following symbols are used: +, binding; NB, no binding; ND, experiment not done.

Dockerin	Native gel	ITC	SPR
			kon	koff	KD
			m−1 s−1	s−1	m
DocScaB	+	+	2.8 × 106	3.5 × 10−5	1.3 × 10−11
DocI15S/N16I	+	+	2.1 × 106	4.2 × 10−5	2.0 × 10−11
DocI51S/N52I	+	+	2.0 × 106	2.9 × 10−5	1.4 × 10−11
DocI15S/N16I/I51S/N52I	NB	NB	NB	NB	NB
DocR14N/K50N	+	+	4.6 × 106	2.9 × 10−4	6.4 × 10−11
DocI15S/I51S	+	+	5.6 × 105	3.1 × 10−4	5.6 × 10−10
DocN16I/N52I	+	+	4.0 × 106	5.4 × 10−2	1.3 × 10−8
DocA18F/A54F	+	+	1.2 × 106	6.7 × 10−2	5.4 × 10−8
DocL20Y/L56Y	+	+	9.1 × 106	2.7 × 10−4	3.0 × 10−11
DocD23Q/D59Q	−	+	1.7 × 106	6.4 × 10−2	3.8 × 10−8
DocGH5	+	+	ND	ND	ND