Fusion maltose-binding protein

With data obtained by the analysis of fusion proteins consisting of a domain unrelated to poly(3HB) metabolism (e.g., maltose binding protein MalE or glutathione-S-transferase and the poly(3HB) depolymerase binding domain [57,59-61]. 

Proteins expressed as fusion proteins have also been crystallized. Maltose binding protein (MBP) (Kobe et al., 1999 Liu et al., 2001), thioredoxin (Stoll et al., 1998) and GST (Kuge et al., 1997) have all been used as fusion proteins in structure studies. 

CitB -I- ATP <121> (<121>, a fusion protein MalE-CitAC is composed of the maltose-binding protein and the CitA kinase domain shows constitutive autokinase activity and transfers the y-phosphate group of ATP to its cognate response regulator CitB [195]) (Reversibility <121> [195]) [195]... 

Riggs, P. (1992) Expression and purification of maltose-binding protein fusions. In Current Protocols in Molecular Biology (Ausubel, F. A., Brent, R., Kingston, R. E., et al., eds.), Greene Publishing and Wiley-Interscience, New York, pp. 16.6.1-16.6.14. 

The His-tagged MBP-MS2 coat fusion protein (HMM) used in this method was created to allow the protein to be immobilized on both Ni2+ or amylose affinity resins. HMM is a 59-kDa protein containing an NT-terminal hexahistidine (6x His) tag, a central maltose-binding protein (MBP) domain, and a C-terminal MS2 coat protein containing the V29/dIFG mutations, which prevent protein multimerization and increase its affinity for RNA (Lim and Peabody, 1994). The protein is expressed in E. coli from plasmid pHMM, which confers kanamycin resistance, and purified using affinity chromatography. 

KD Pryor, B Leiting. High-level expression of soluble protein in Escherichia coli using a His6-tag and maltose-binding protein double-affinity fusion system. Prot Expr Purif 10 309-319, 1997. 

Riggs, P. (2000) Expression and purification of recombinant proteins by fusion to maltose-binding protein. Mol. Biotechnol. 15, 51-63. 

In this work, a fusion protein has been engineered from maltose binding protein (pmal) and human metallothionein (MT). The fusion protein (pmal-MT) has been expressed in E. coli, and purified with an amylose column. The purified fusion protein was immobilized on a solid matrix, and its characteristics as metal binding ligand have been studied. We have found that the pmal-MT ligand efficiently binds gallium ion, one of the valuable rare metals desired to be recovered from aqueous solution [3]. Different binding mechanisms for two metal ions have been elucidated based on HSAB (hard and soft acids and bases) theory [4]. 

Weller U, Moller L, Mefiner M et at. (1996) Expression of active streptolysin-O in E. coti as maltose-binding protein-streptolysin-O fusion protein The N-terminal 70 amino acids are not required for hemolytic activity. Eur. J. Biochem. 236 34-39... 

The simplest modification is to place a destabilizing amino acid at the N-terminal end according to the N-end rule (Varshavsky, 1992). This can be done by expressing the protein as a fusion protein, e.g. with maltose binding protein behind a cleavage site specific for a rare-cutting protease, such as factor Xq. In this case the N-terminal amino acid can be any amino acid except proline. 

DNA sequence determination and analysis [22] functional expression in Escherichia coli strain BL21(DE3) as His-tagged protein [22,25]) [22, 25] <14> (expression of truncated enzyme forms, comprising residues 284-402 and 1-334, respectively, as maltose-binding-protein fusion proteins in Escherichia coli JM109 [25] functional expression in Escherichia coli as His-tagged protein [25]) [25]... 

Bassford P J, Silhavy T J, Beckwith J R (1979). Use of gene fusion to study secretion of maltose-binding protein into Escherichia coli periplasm. J. Bact. 139 19-31. Yamanaka H, Nomura T, Fuji Y, Okamoto K (1998). Need for TolC, an Escherichia coli outer membrane protein, in the secretion of heat-stable enterotoxin I across the outer membrane. Microb. Pathogen. 25 111-120. 

Toda A, Yamada K, Nishimura S-I. An engineered biocatalyst for the synthesis of glycoconjugates utilization of (31,3-V-acetyl-D-glucosaminyltransferase from Streptococcus agalactiae type la expressed in Escherichia coli as a fusion with maltose-binding protein. Adv Synth Catal 2002 344 61-69. 

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