Display of a functional hetero-oligomeric catalytic antibody on the yeast cell surface.
A functional hetero-oligomeric protein was, for the first time, displayed on the yeast cell surface. A hetero-oligomeric Fab fragment of the catalytic antibody 6D9 can hydrolyze a non-bioactive chloramphenicol monoester derivative to produce chloramphenicol. The gene encoding the light chain of the Fab fragment of 6D9 was expressed with the tandemly-linked C-terminal half of alpha-agglutinin. At the same time, the gene encoding the Fd fragment of the heavy chain of the Fab fragment was expressed as a secretion protein. The combined Fab fragment displayed and associated on the yeast cell surface had an intermolecular disulfide linkage between the light and heavy chains. This protein fragment catalyzed the hydrolysis of a chloramphenicol monoester derivative and exhibited high stability in binding with a transition-state analog (TSA). The catalytic reaction was also inhibited by the TSA. The successful display of a functional hetero-oligomeric catalytic antibody provides a useful model for the display of hetero-oligomeric proteins and enzymes.
Lin
Y
Y
Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, 606-8501, Kyoto, Japan.
Tsumuraya
T
T
Wakabayashi
T
T
Shiraga
S
S
Fujii
I
I
Kondo
A
A
Ueda
M
M
D017151:Antibodies, Catalytic / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D002462:Cell Membrane / Q000276:immunology; D002701:Chloramphenicol / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D007700:Kinetics; D019151:Peptide Library; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000276:immunology*
D017151:Antibodies, Catalytic; D004274:DNA, Recombinant; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011994:Recombinant Proteins; D002701:Chloramphenicol