Functional and structural characterization of chimeras of a bacterial genotoxin and human type I DNAse.
Chimeras composed of the cdtB gene of a novel bacterial genotoxin and the human type I DNAse I gene were constructed and their products characterized relative to the biochemical and enzymatic properties of the native proteins. The product of a cdtB/DNAse I chimera formed a heterotrimer with the CdtA and CdtC subunits of the genotoxin, and targeted mutations increased the specific activity of the hybrid protein. Expression of active chimeric gene products established that the CdtB protein is an atypical divalent cation-dependent endonuclease and demonstrated the potential for genetically engineering a new class of therapeutic agent for inhibiting the proliferation of cancer cells.
DiRienzo
Joseph M
JM
Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104-6030, USA. dirienzo@pobox.upenn.edu
Cao
Linsen
L
Volgina
Alla
A
Bandelac
Georges
G
Korostoff
Jonathan
J
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001427:Bacterial Toxins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003850:Deoxyribonuclease I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D015661:Pasteurellaceae / Q000378:metabolism*; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D011993:Recombinant Fusion Proteins; C067044:cytolethal distending toxin; D003850:Deoxyribonuclease I