ISSN:
1432-1327
Keywords:
Key words Human immunodeficiency virus type 1 reverse transcriptase ribonuclease H domain
;
Calorimetry
;
Metal binding
;
Mechanism
;
Stoichiometry
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
Abstract Crystallographic studies of the Mn2+-doped RNase H domain of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) [1] have revealed two bound Mn2+ separated by approximately 4 Å and surrounded by a cluster of four conserved carboxylates. Escherichia coli RNase H is structurally similar to the RNase H domain of HIV-1 RT, but requires one divalent metal cation for its activity [2, 3], implying either that the HIV-1 RT RNase H domain contrasts in its ability to bind two divalent metal ions, or that the crystallographic data reflect specific use of Mn2+ and/or the doping technique employed. Metal binding stoichiometry has been determined for Mn2+ and the biologically more relevant Mg2+ cation by solution calorimetric studies of native and recombinant p66/p51 HIV-1 RT. Three Mn2+ ions bind to HIV-1 RT apo-enzyme: one at the DNA polymerase and two at the RNase H catalytic center, the latter being consistent with crystallographic results. However, only one Mg2+ ion is bound in the RNase H catalytic center. Several mechanistic implications arise from these results, including the possibility of mutually exclusive Mg2+ binding sites that might be occupied according to the specific reaction being catalyzed by the multifunctional RNase H domain. The occurrence of distinct binding stoichiometries for Mg2+ and Mn2+ to multifunctional enzymes has previously been reported [4].
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s007750050009
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