Divalent metal cation binding properties of human prothymosin alpha.
The divalent cation binding properties of human prothymosin alpha, an abundant nuclear protein involved in cell proliferation, were evaluated. By using prothymosin alpha retardation on a weak cation chelating resin charged with various divalent cations, specific binding of Zn2+ ions by prothymosin alpha was observed. This finding was further confirmed by the equilibrium dialysis analysis which demonstrated that, within the micromolar range of Zn2+ concentrations, prothymosin alpha could bind up to three zinc ions in the presence of 100 mM NaCl and up to 13 zinc ions in the absence of NaCl. Equilibrium dialysis analysis also revealed that prothymosin alpha could bind Ca2+, although the parameters of Ca2+ binding by prothymosin alpha were less pronounced than those of Zn2+ binding in terms of the number of metal ions bound, the KD values, and the resistance of the bound metal ions to 100 mM NaCl. The effects of Zn2+ and Ca2+ on the interaction of prothymosin alpha with its putative partners, Rev of HIV type 1 and histone H1, were examined. We demonstrated that Rev binds prothymosin alpha, and that prothymosin alpha binding to Rev but not to histone H1 was significantly enhanced in the presence of zinc and calcium ions. Our data suggest that the modes of prothymosin alpha interaction with Rev and histone H1 are distinct and that the observed zinc and calcium-binding properties of prothymosin alpha might be functionally relevant.
Chichkova
N V
NV
Belozersky Institute of Physico-Chemical Biology and Center of Molecular Medicine, Moscow State University, Russia.
Evstafieva
A G
AG
Lyakhov
I G
IG
Tsvetkov
A S
AS
Smirnova
T A
TA
Karapetian
R N
RN
Karger
E M
EM
Vartapetian
A B
AB
D002118:Calcium / Q000378:metabolism; D002412:Cations; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D015694:Gene Products, rev / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011498:Protein Precursors / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013947:Thymosin / Q000031:analogs & derivatives* / Q000378:metabolism; D013997:Time Factors; D015032:Zinc / Q000378:metabolism; D054321:rev Gene Products, Human Immunodeficiency Virus
D002412:Cations; D015694:Gene Products, rev; D006657:Histones; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C040782:prothymosin alpha; D054321:rev Gene Products, Human Immunodeficiency Virus; D013947:Thymosin; D008274:Magnesium; D015032:Zinc; D002118:Calcium