Differential degradation of a recombinant albumin-binding receptor in Escherichia coli.
The degradation in Escherichia coli of the recombinant serum-albumin-binding receptor derived from streptococcal protein G was investigated using a dual-affinity fusion approach. The proteolytic degradation of the receptor was characterized when fused to human proinsulin and human secretin. Several cleavages occurred at sequences not normally regarded as proteolytically sensitive, such as the dipeptide sequences Ile-Gly, Val-Ser and Ser-Ala. Depending on the fusion partner, large differences in the degradation of the albumin-binding domain were observed. Thus, susceptibility to proteolysis of a recombinant protein can be affected by a neighbouring domain.
Murby
M
M
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Nygren
P A
PA
Rondahl
H
H
Hellman
U
U
Enfors
S O
SO
Uhlén
M
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D011384:Proinsulin / Q000096:biosynthesis / Q000302:isolation & purification; D017457:Receptors, Albumin; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012633:Secretin / Q000096:biosynthesis / Q000302:isolation & purification; D012709:Serum Albumin / Q000378:metabolism*
D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D017457:Receptors, Albumin; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D012709:Serum Albumin; D012633:Secretin; D011384:Proinsulin