Design of a serine protease-like catalytic triad on an antibody light chain displayed on the yeast cell surface

  title={Design of a serine protease-like catalytic triad on an antibody light chain displayed on the yeast cell surface},
  author={Norihiko Okochi and Michiko Kato-Murai and Tetsuya Kadonosono and Mitsuyoshi Ueda},
  journal={Applied Microbiology and Biotechnology},
Lc-WT, the wild-type light chain of antibody, and Lc-Triad, its double mutant with E1D and T27aS designing for the construction of catalytic triad within Asp1, Ser27a, and original His93 residues, were displayed on the cell surface of the protease-deficient yeast strain BJ2168. When each cell suspension was reacted with BODIPY FL casein and seven kinds of peptide-MCA substrates, respectively, a remarkable difference in hydrolytic activities toward Suc-GPLGP-MCA (succinyl-Gly-Pro-Leu-Gly-Pro-MCA… 
Finding and characterizing a catalytic antibody light chain, H34, capable of degrading the PD-1 molecule
A unique human catalytic antibody light chain, H34, is described, which mediates enzymatic degradation of human PD-1 peptides and recombinant humanPD-1 protein and thus functions to prevent the binding of PD- 1 with PD-L1.
Expression of catalytic antibodies in eukaryotic systems
Calculated values indicate that activity of the antibodies expressed in yeast is similar to the full-size antibody A17 and to the single-chain antibody A.17 expressed in CHO and E. coli cells, respectively.
Biochemical Features of a Catalytic Antibody Light Chain, 22F6, Prepared from Human Lymphocytes*
The 22F6 catalytic light chain possesses two kinds of active sites as amidase and nuclease in close distances and could suppress the infection of influenza virus type A (H1N1) of Madin-Darby canine kidney cells in an in vitro assay.
New technologies to introduce a catalytic function into antibodies: A unique human catalytic antibody light chain showing degradation of β‐amyloid molecule along with the peptidase activity
This study screened a protein bank of human antibody light chains, in which the light chains were expressed, purified, and stored for use in screening against many kinds of antigen, to get clues to introducing a catalytic function in normal antibodies.
Alteration of substrate specificity of rat neurolysin from matrix metalloproteinase-2/9-type to -3-type specificity by comprehensive mutation.
Results indicate that Tyr610 of neurolysin is the important residue to recognize the P2' amino acid of substrates.
Highly efficient method of preparing human catalytic antibody light chains and their biological characteristics
It is demonstrated that high‐throughput selection of light chains possessing catalytic functions and specificity for a target molecule can be attained from a light‐chain DNA library amplified from germline genes belonging to subgroup II.
[Novel high-throughput system for production of new medicines-integration and combination with molecular display and combinatorial bioengineering].
  • M. Ueda
  • Biology, Chemistry
    Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan
  • 2009
To demonstrate the practical use of a novel high-throughput screening system by single cells constructed by the molecular display method, a yeast cell chip microchamber array was developed. As
A Monoclonal Antibody to Cryptococcus neoformans Glucuronoxylomannan Manifests Hydrolytic Activity for Both Peptides and Polysaccharides*
The ability of many antibodies to cleave antigen, albeit slowly, supports the notion that this activity is an important immunoglobulin function in host defense and suggests new therapeutic possibilities for polysaccharide-binding antibodies.
Development of a yeast protein fragment complementation assay (PCA) system using dihydrofolate reductase (DHFR) with specific additives
Demonstrations suggest that the yeast PCA system based on DHFR can be one of good, convenient, and inexpensive tools for investigating eukaryotic protein–protein interactions in vivo.
Yeast Cell Surface Display of Bacterial Chitinase as a New Approach for Biocontrol of Phytopathogenic Fungi
It is demonstrated that chitinase was successfully displayed on the yeast cell surface in an active form with an antifungal activity against phytopathogenic fungi, especially Fusarium oxysporum.


Site-directed mutagenesis of proteolytic antibody light chain.
Two types of residues participating in catalysis by the light chain have been identified: Ser27a and His93 are essential for catalysis but not for initial high affinity complexation and substrate, and Ser26 and His27d or Asp28 participate in VIP binding and limit turnover indirectly.
Display of a functional hetero-oligomeric catalytic antibody on the yeast cell surface
The successful display of a functional hetero-oligomeric catalytic antibody provides a useful model for the display of hetero,oligomersic proteins and enzymes.
Cleavage specificity of a proteolytic antibody light chain and effects of the heavy chain variable domain.
A model of catalysis by the anti-VIP Fv is led in which the essential catalytic residues are located in the VL domain and additional residues from the VH domain are involved in high affinity binding of the substrate.
Catalytic antibody light chain capable of cleaving a chemokine receptor CCR‐5 peptide with a high reaction rate constant
A monoclonal antibody (MAb), ECL2B‐2, was obtained by immunizing a peptide possessing a part of a sequence of a chemokine receptor, CCR‐5, which is present as a membrane protein on the macrophage
A mutagenesis study of a catalytic antibody.
Results not only demonstrate the importance of electrostatic interactions in catalysis by antibody S107 but also show that catalytic side chains can be introduced into antibodies to enhance their catalytic efficiency.
Endopeptidase character of monoclonal antibody i41-7 subunits.
Generation of a catalytic antibody by site-directed mutagenesis.
A hybrid Fv fragment of the dinitrophenyl-binding immunoglobulin A (IgA), MPOC315, has been generated by reconstituting a recombinant variable light chain (VL) produced in Escherichia coli with a
Natural Catalytic Antibodies: Peptide-hydrolyzing Activities of Bence Jones Proteins and VL Fragment (*)
Results provide evidence for the proteolytic activity of certain human light chains and imply that this phenomenon may have a pathophysiological significance.
Analysis of a processing system for proteases using yeast cell surface engineering: conversion of precursor of proteinase A to active proteinase A
It was suggested that by using the yeast-cell-surface engineering at the location of the initial step, the autocatalytic activation from proPrA to PrA might occur before the vacuolar branch separates from the main secretory pathway.
Molecular cloning of a proteolytic antibody light chain.