p10 single-stranded nucleic acid binding protein from murine leukemia virus binds metal ions via the peptide sequence Cys26-X2-Cys29-X4-His34-X4-Cys39.
The RNA binding protein of 56 residues encoded by the extreme 3' region of the gag gene of Rauscher murine leukemia virus (MuLV) has been chemically synthesized by a solid-phase synthesis approach. Since the peptide contains a Cys26-X2-Cys29-X4-His34-X2-Cys39 sequence that is shared by all retroviral gag polyproteins which has been proposed to be a metal binding region, it was of considerable interest to examine the metal binding properties of the complete p10 protein. As postulated, p10 binds the metal ions Cd(II), Co(II), and Zn(II). The Co(II) protein shows a set of d-d absorption bands typical of a tetrahedral Co(II) complex at 695 (epsilon = 565 M-1 cm-1), 642 (epsilon = 655 M-1 cm-1), and 615 nm (epsilon = 510 M-1 cm-1) and two intense bands at 349 (epsilon = 2460 M-1 cm-1) and 314 nm (epsilon = 4240 M-1 cm-1) typical of Co(II)----(-)S- charge transfer. The ultraviolet absorption spectrum also indicates Cd(II) binding by the appearance of a Cd(II)----(-)S- charge-transfer band at 255 nm. The 113Cd NMR spectrum of 113Cd(II)-p10 reveals one signal at delta = 648 ppm. This chemical shift correlates well with that predicted for ligation of 113Cd(II) to three -S- from the three Cys residues of p10. The chemical shift of 113Cd(II)-p10 changes by only 4 ppm upon binding of d(pA)6, indicating that the chelate complex is little changed by oligonucleotide binding.(ABSTRACT TRUNCATED AT 250 WORDS)
Roberts
W J
WJ
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06510.
Pan
T
T
Elliott
J I
JI
Coleman
J E
JE
Williams
K R
KR
D000595:Amino Acid Sequence; D002104:Cadmium / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D002942:Circular Dichroism; D003035:Cobalt / Q000378:metabolism*; D015683:Gene Products, gag / Q000378:metabolism; D007554:Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D011485:Protein Binding; D011925:Rauscher Virus / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism; D015032:Zinc / Q000378:metabolism*
D002352:Carrier Proteins; D015683:Gene Products, gag; D007554:Isotopes; D014764:Viral Proteins; D002104:Cadmium; D003035:Cobalt; D015032:Zinc