A synthetic miniprotein that binds specific DNA sequences by contacting both the major and the minor groove.
Attachment of a slightly modified basic region of a bZIP protein (GCN4) to a distamycin-related tripyrrole provides a bivalent system capable of binding with high affinity to specific DNA sequences. Appropriate adjustment of the linker between the two units has led to a hybrid that binds a 9 base-pair-long DNA site (TTTTATGAC) with low nanomolar affinity at 4 degrees C. Circular dichroism and gel retardation studies indicate that the binding occurs by simultaneous insertion of the bZIP basic region into the DNA major groove and the tripyrrole moiety into the minor groove of the flanking sequence. Analysis of hybrids bearing alternative linkers revealed that tight, specific binding is strongly dependent on the length and nature of the connecting unit.
Blanco
Juan B
JB
Departamento de Química Orgánica y, Unidad Asociada al CSIC, Universidad de Santiago de Compostela, 15782, Santiago de Compostela, Spain.
Vázquez
M Eugenio
ME
Martinez-Costas
José
J
Castedo
Luis
L
Mascareñas
José L
JL
D000595:Amino Acid Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D004214:Distamycins / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011494:Protein Kinases / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D011758:Pyrroles / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship
D004268:DNA-Binding Proteins; D004214:Distamycins; D011758:Pyrroles; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C030000:stallimycin; D004247:DNA; D011494:Protein Kinases