Investigation of active form of catalytic antibody light chain 41S-2-L

Mitsuda, Yukie; Tsuruhata, Kumi; Hifumi, Emi; Takagi, Masahiro; Uda, Takaaki

doi:10.1016/j.imlet.2004.07.018

Cited by 9 publications

(18 citation statements)

References 21 publications

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“…In this regard, variations in the hydrolytic activity and nucleophilic properties were found between different A18b preparations; however, these could be restored to similar levels through denaturation and refolding. Indeed, light-chain dimers can form multiple structures under different crystallization conditions (36), and aggregates have been proposed to function in the catalytic mechanism (37). The relationship between the biophysical characteristics of various light chains and myeloma pathogenesis is not yet clear.…”

Section: Discussionmentioning

confidence: 99%

A Human Germ Line Antibody Light Chain with Hydrolytic Properties Associated with Multimerization Status

Sharma¹,

Heriot²,

Trisler³

et al. 2009

Journal of Biological Chemistry

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Antibodies with nucleophilic or catalytic properties often have these characteristics encoded in their germ line genes. Because hydrolytic activity has been reported to be associated with light chain V regions, we have begun an analysis of germ line light chain proteins that could be the basis for affinity maturation into hydrolytic or other reactive antibodies. We produced the germ line A18b light chain and characterized its hydrolytic, nucleophilic, and tertiary structural activities. This light chain was purified to >99% purity and found to hydrolyze aminomethylcoumarin-peptide and larger protein substrates and bind a fluorophosphonate probe. Mutation of putative catalytic residues only resulted in loss of activity of a tetrameric but not dimeric form of the light chain. These biochemical properties provide a framework for understanding the structure-function relationships of germ line antibodies.Antibodies are responsible for defense against invading pathogens, but they can also be pathogenic in autoimmune disease. Antibody genes are partially hard-coded in the genome as gene modules known as V, D, and J segments for the heavy chain, or V and J segments for the light chain (1). Multiple modules allow for the diversity of the antibody repertoire through combinatorial rearrangement. During B-cell development, the germ line-encoded V, D, and J segments recombine to form functional heavy-and light-chain V region genes. Following exposure to antigen, B-cells activate an affinity maturation process that mutates V regions, and those B-cells with higher affinity variants are driven to survive and proliferate. Typically, germ line-derived antibodies have affinities in the mid nanomolar to micromolar range (2, 3), as opposed to nanomolar or less for affinity-matured antibodies. Germ line-derived antibodies contain unique properties, including polyspecificity and catalytic activity. The polyspecificity is thought to be due to an inherent flexibility in germ line antibody complementarity-determining regions (CDRs), 2 which allows them to adopt multiple conformations and bind different antigens (4 -6). The structural basis for catalytic activity is more obscure, however, structural studies of induced catalytic antibodies also show multiple structures in their germ line precursors (4,7,8).Recently, catalytic antibodies have been associated with positive or negative aspects of human disease (9, 10). In sepsis, catalytic antibodies were predictive of survival (11). In subsets of hemophilia, proteolytic antibodies against Factor VIII were associated with resistance to therapy (12, 13). Catalytic antibodies against myelin basic protein in multiple sclerosis have also been found and suggested to contribute to pathogenesis (14, 15). In multiple myeloma, Bence-Jones light-chain proteins can contribute to pathogenesis of the disease and in some cases have been reported to contain catalytic activity (16,17). The structural and biochemical details around such antibodies have not yet been studied in detail.We describe here a human g...

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Section: Discussionmentioning

confidence: 99%

A Human Germ Line Antibody Light Chain with Hydrolytic Properties Associated with Multimerization Status

Sharma¹,

Heriot²,

Trisler³

et al. 2009

Journal of Biological Chemistry

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“…Since the original discovery of autoantibody-mediated antigen degradation [13,16], a wide spectrum of the disease-related abzymes has been described [17][18][19][20][21]. Unfortunately, the role of the antibody-mediated catalysis in the pathogenesis of autoimmune and other disorders is mostly unclear.…”

Section: Discussionmentioning

confidence: 99%

Catalytic activity of autoantibodies toward myelin basic protein correlates with the scores on the multiple sclerosis expanded disability status scale

Пономаренко

Durova²,

Vorobiev

et al. 2006

Immunology Letters

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“…As a result, three amino acids (Ser, His, and Asp) can come into a close position so as to make a catalytic triad showing the catalytic activity. We have already reported that the conformational change of the light chain takes place in the induction period of the cleavage reaction (12). During the cleavage reaction, the enzymatic activity of urease declined to 3%, indicating that UA15-L destroyed not only the ␤-subunit but also the enzymatic function of the urease.…”

Section: Discussionmentioning

confidence: 93%

“…Cleavage Activity against UreB-Because the epitope of UA15 mAb is not determined, a peptidase activity of UA15-L was examined by using a TP41-1 peptide (TPRGPDRPEGIEEE-GGERDRD), which has been used to monitor the peptidase activity of the catalytic antibody or its subunits (3,12,13,15,16). The degradation of TP41-1 peptide by UA15-L followed the Michaelis-Menten equation, and its kinetic parameters were obtained as k cat ϭ 0.24 min Ϫ1 and K m ϭ 6.2 ϫ 10 Ϫ5 M (see supplemental Appendix S1).…”

Section: Resultsmentioning

confidence: 99%

“…Prior to the cleavage test for UreB protein and the purified urease of H. pylori, a peptide, TPRGPDRPEGIEEEGGERDRD (TP41-1), which has mostly been used for monitoring the catalytic activity of the antibody and/or its subunits (3,10,(12)(13)(14), was completely digested by the catalytic reaction of UA15-L. By this reaction, the catalytic activity of UA15-L was held constant, showing no induction time (3,(12)(13)(14)(15). In the cleavage assay of UreB and the urease, the UA15-L mentioned above was used.…”

Section: Cleavage Assaysmentioning

confidence: 99%

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Catalytic Features and Eradication Ability of Antibody Light-chain UA15-L against Helicobacter pylori

Hifumi

Morihara²,

Hatiuchi

et al. 2008

Journal of Biological Chemistry

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We have successfully developed a catalytic antibody capable of degrading the active site of the urease of Helicobacter pylori and eradicating the bacterial infection in a mouse stomach. This monoclonal antibody UA15 was generated using a designed recombinant protein UreB, which contained the crucial region of the H. pylori urease ␤-subunit active site, for immunization. The light chain of this antibody (UA15-L) by itself showed a proteolytic activity to substantially degrade both UreB and the intact urease. Oral administration of UA15-L also significantly reduced the number of H. pylori in a mouse stomach. This is the first example of a monoclonal catalytic antibody capable of functioning in vivo, and such an antibody may have a therapeutic utility in the future.

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Investigation of active form of catalytic antibody light chain 41S-2-L

Cited by 9 publications

References 21 publications

A Human Germ Line Antibody Light Chain with Hydrolytic Properties Associated with Multimerization Status

A Human Germ Line Antibody Light Chain with Hydrolytic Properties Associated with Multimerization Status

Catalytic activity of autoantibodies toward myelin basic protein correlates with the scores on the multiple sclerosis expanded disability status scale

Catalytic Features and Eradication Ability of Antibody Light-chain UA15-L against Helicobacter pylori

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