Abstract
We have constructed an allosterically controllable novel enzyme (designated maxizyme) that can be transcribed in vivo under the control of a human tRNA(Val) promoter. The maxizyme has sensor arms that can recognize target sequences, and in the presence of such a target sequence only, it can form a cavity that can capture catalytically indispensable Mg2+ ions. As a target for a demonstration of the potential utility of the maxizyme, we chose BCR-ABL mRNA, the translated products of which cause chronic myelogenous leukemia. Only the maxizyme (but not conventional ribozymes) had extremely high specificity and high-level activity, not only in vitro but also in cultured cells including BV173 cells derived from a patient with a Philadelphia chromosome. The maxizyme induced apoptosis only in leukemic cells with this chromosome.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Allosteric Regulation
-
Animals
-
Apoptosis / drug effects
-
Base Sequence
-
Caspase 3
-
Caspases / metabolism
-
Cell Line
-
Cell Line, Transformed
-
Cytoplasm / metabolism
-
Dimerization
-
Enzyme Precursors / metabolism
-
Fusion Proteins, bcr-abl / genetics*
-
Gene Expression
-
Humans
-
Interleukin-3 / pharmacology
-
Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics
-
Leukemia, Myelogenous, Chronic, BCR-ABL Positive / metabolism*
-
Leukemia, Myelogenous, Chronic, BCR-ABL Positive / therapy
-
Magnesium / pharmacology
-
Mice
-
Nucleic Acid Conformation
-
Philadelphia Chromosome
-
RNA, Catalytic / metabolism*
-
RNA, Catalytic / therapeutic use
-
RNA, Messenger / metabolism*
-
RNA, Transfer, Val / genetics
-
Substrate Specificity
-
Transfection
-
Tumor Cells, Cultured
Substances
-
Enzyme Precursors
-
Interleukin-3
-
RNA, Catalytic
-
RNA, Messenger
-
RNA, Transfer, Val
-
Fusion Proteins, bcr-abl
-
CASP3 protein, human
-
Casp3 protein, mouse
-
Caspase 3
-
Caspases
-
Magnesium