HUMAN RECONSTITUTED DNA TOPOISOMERASE I IN COVALENT COMPLEX WITH A 22 BASE PAIR DNA DUPLEX

Crystal structures of human topoisomerase I in covalent and noncovalent complexes with DNA.

DNA (5'-D(*AP*AP*AP*AP*AP*GP*AP*CP*5IUP*5IU*TP*GP*AP*AP*AP*AP*AP*5IUP*5IUP*5IUP*5IUP*T)-3')

DNA (5'-D(*AP*AP*AP*AP*AP*TP*5IUP*5IUP*5IUP*5IUP*CP*AP*AP*AP*GP*TP*CP*TP*TP*TP*TP*T)-3')

KPKNKDKDKKVPEPDNKKKKPKKEEEQKWKWWEEERYPEGIKWKFLEHKGPVFAPPYEPLPENVKFYYDGKVMKLSPKAE
EVATFFAKMLDHEYTTKEIFRKNFFKDWRKEMTNEEKNIITNLSKCDFTQMSQYFKAQTEARKQMSKEEKLKIKEENEKL
LKEYGFCIMDNHKERIANFKIEPPGLFRGRGNHPKMGMLKRRIMPEDIIINCSKDAKVPSPPPGHKWKEVRHDNKVTWLV
SWTENIQGSIKYIMLNPSSRIKGEKDWQKYETARRLKKCVDKIRNQYREDWKSKEMKVRQRAVALYFIDKLALRAGNEKE
EGETADTVGCCSLRVEHINLHPELDGQEYVVEFDFLGKDSIRYYNKVPVEKRVFKNLQLFMENKQPEDDLFDRLNTGILN
KHLQDLMEGLTAKVFRTYNASITLQQQLKELTAPDENIPAKILSYNRANRAVAILCNHQRAPPKTFEKSMMNLQTKIDAK
KEQLADARRDLKSAKADAKVMKDAKTKKVVESKKKAVQRLEEQLMKLEVQATDREENKQIALGTSKLN(PTR)LDPRITV
AWCKKWGVPIEKIYNKTQREKFAWAIDMADEDYEF

KPKNKDKDKKVPEPDNKKKKPKKEEEQKWKWWEEERYPEGIKWKFLEHKGPVFAPPYEPLPENVKFYYDGKVMKLSPKAE
EVATFFAKMLDHEYTTKEIFRKNFFKDWRKEMTNEEKNIITNLSKCDFTQMSQYFKAQTEARKQMSKEEKLKIKEENEKL
LKEYGFCIMDNHKERIANFKIEPPGLFRGRGNHPKMGMLKRRIMPEDIIINCSKDAKVPSPPPGHKWKEVRHDNKVTWLV
SWTENIQGSIKYIMLNPSSRIKGEKDWQKYETARRLKKCVDKIRNQYREDWKSKEMKVRQRAVALYFIDKLALRAGNEKE
EGETADTVGCCSLRVEHINLHPELDGQEYVVEFDFLGKDSIRYYNKVPVEKRVFKNLQLFMENKQPEDDLFDRLNTGILN
KHLQDLMEGLTAKVFRTYNASITLQQQLKELTAPDENIPAKILSYNRANRAVAILCNHQRAPPKTFEKSMMNLQTKIDAK
KEQLADARRDLKSAKADAKVMKDAKTKKVVESKKKAVQRLEEQLMKLEVQATDREENKQIALGTSKLNYLDPRITVAWCK
KWGVPIEKIYNKTQREKFAWAIDMADEDYEF

pH 7.7, VAPOR DIFFUSION, SITTING DROP, temperature 295.00K

RESIDUES 215-635 OF CHAIN A COMPRISE THE CORE DOMAIN OF
THE MOLECULE. RESIDUES 715-765 OF CHAIN A COMPRISE THE
C-TERMINAL DOMAIN. 1-22 OF CHAIN C COMPRISE THE SCISSILE
STRAND OF THE DNA, 101-122 OF CHAIN D THE INTACT DNA
STRAND.

NOTE THAT THE LINKER DOMAIN (636-714) IS NOT
PRESENT IN THE CONSTRUCT OF THIS RECONSTITUTED ENZYME.
THE RECONSTITUTED ENZYME IS MADE UP OF RESIDUES 175-659 OF
THE CORE (EXPRESSED IN BACULOVIRUS) AND RESIDUES 713-765 OF
THE C-TERMINAL DOMAIN (EXPRESSED IN E. COLI).

CHAIN A RESIDUES 175-214 AND 627-659 OF THE CORE AND
713-719 OF THE C-TERMINAL DOMAIN ARE DISORDERED AND NOT
PRESENT IN THE STRUCTURE.

TER
ALA 635 A IS THE TERMINAL RESIDUE OF THE CORE DOMAIN.
PHE 765 A IS THE TERMINAL RESIDUE OF THE C-TERMINAL DOMAIN.
THY 22 C IS THE 3' TERMINAL NUCLEOTIDE OF THE SCISSILE
STRAND OF THE DNA DUPLEX.
THY 122 D IS THE 3' TERMINAL NUCLEOTIDE OF THE INTACT
STRAND OF THE DNA DUPLEX.

MULTIPLE ISOMORPHOUS REPLACEMENT WITH ANOMALOUS SCATTERING

TOPOISOMERASE I (E.C.5.99.1.2)/DNA COMPLEX

TOPOISOMERASE I/DNA, DNA, TOPOISOMERASE I, ISOMERASE-DNA COMPLEX