Refolding Record:

Protein See all refolding records for this protein...
Protein Name	Glyceraldehyde-3-phosphate dehydrogenase
Abbreviated Name	GAPDH
SCOP Family	Unknown
Structure Notes
Organism	Streptomyces avidinii
UniProt Accession	P22629
SCOP Unique ID	n/a
Structure Solved
Class	Unknown
Molecularity	Tetramer

Construct
Full Length	y
Domain	n/a
Chimera	n/a
Variants	n/a
Chain Length	159
Molecular Weight	16490.9
Pi	6.1
Molecular Weight	16490.9
Disulphides	Unknown
Full Sequence	DPSKDSKAQVSAAEAGITGTWYNQLGSTFIVTAGADGALTGTYESAVGNAESRYVLTGRYDSAPATDGSGTALGWTVAWKNNYRNAHSATTWSGQYVGGAEARINTQWLLTSGTTEANAWKSTLVGHDTFTKVKPSAASIDAAKKAGVNNGNPLDAVQQ
Notes	n/a

Expression
Report	Liu X, Liu J. (2005) Biotechnol Lett, 27, 1067-1073
Project Aim	Undefined
Fusion	None
Protein Expression and Production	Protein recombinantly expressed as and refolded from inclusion bodies.
Expression Host	Escherichia coli
Expression Strain	BL21(DE3)
Expression Temp	0.0
Expression Time	00
Expression Vector	pET28a
Expression Protocol	Expression and purification of streptavidins Three recombinant streptavidins (rsavC, rsavF and rsavS) were expressed and purified using a Ni2+ affinity column according to the manufacturer’s protocol (Novagen). In order to evaluate the ratio of soluble streptavidin to insoluble protein expressed as inclusion bodies and to establish the best protocol for production of active streptavidin, three streptavidins were purified under both native and denaturing conditions. 15% (w/v) SDS- PAGE was used to evaluate the purity of purified proteins. Protein concentrations were determined by Bradford’s method.
Method of Induction	Not Stated
Cell Density at Induction	OD n/a = n/a
Cell Disruption Method	Not stated
Lytic Agent	None
Pre-Refolding Purification	Metal affinity chromatography
Solubility	insoluble

Refolding
Refolding Method	Dialysis
Wash Buffer	n/a
Solubilization Buffer	n/a
Refolding Buffer	20 mM Tris/HCl pH 7.4, 20 mM NaCl and 5 mM B-mercaptoethanol
Pre-Refolding Purification	Metal affinity chromatography
Tag Cleaved	no tag
Refolding pH	7.4
Refolding Temperature	25.0
Protein Concentration	n/a
Refolding Time	n/a
Redox Agent	Beta-mercaptoethanol
Redox Agent Concentration	5 mM
Refolding Protocol	Refolding of denaturing-purified proteins Denaturing-purified streptavidins (0.5 mg/ml) were successively dialyzed against refolding buffer (20 mM Tris/HCl pH 7.4, 20 mM NaCl and 5 mM B-mercaptoethanol) containing decreasing concentrations of urea (4, 2, 1, 0 M). Dialysis was performed for 6 h at each urea level. During dialysis, 50 ll aliquots was taken out at each urea level and centrifuged at 12 000 g for 20 min for determining protein concentrations in the supernatants by Bradford’s method, which indicate the renaturing efficiency of denatured protein.
Refolding Assay	Unspecified
Refolding Chaperones	None
Refolding Additives	None
Additives Concentration	n/a
Refolding Yield	n/a
Purity	n/a
Notes	n/a

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