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Catalytic digestion of human tumor necrosis factor‐α by antibody heavy chain
Author(s) -
Hifumi Emi,
Higashi Kyohei,
Uda Taizo
Publication year - 2010
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2010.07785.x
Subject(s) - chemistry , tumor necrosis factor alpha , antibody , immunoglobulin light chain , cytokine , protease , monoclonal antibody , microbiology and biotechnology , cleavage (geology) , biochemistry , immunology , biology , enzyme , paleontology , fracture (geology)
It has long been an important task to prepare a catalytic antibody capable of digesting a targeting crucial protein that controls specific life functions. Tumor necrosis factor‐α (TNF‐α) is a cytokine and an important molecule concerned with autoimmune diseases such as rheumatoid arthritis, chronic obstructive pulmonary disease, and Crohn’s disease. A mAb (ETNF‐6 mAb) raised against human TNF‐α was prepared, and the steric conformation was created by using molecular modeling after the cDNA was sequenced. The heavy chain (ETNF‐6‐H) of the mAb was considered to possess a catalytic triad‐like structure in the complementarity determining regions (CDRs). As a result, ETNF‐6‐H exhibited a peptidase and a protease activity. In fact, ETNF‐6‐H predominantly cleaved the Ser5‐Arg6 bond of TNF‐α at the first step, resulting in the generation of a fragment of ∼ 17 kDa. This fragment was digested to a smaller molecule of 15 kDa by scission of the Gln21‐Ala22 bond. The intermediate product was further converted into a fragment of 13.3 kDa by successive cleavage of the Leu36‐Leu37 and Asn39‐Gly40 bonds. The heavy chain possessed a protease activity against TNF‐α with a multicleavage site.