N-terminal fragment of the anti-angiogenic human endostatin binds copper(II) with very high affinity.
A histidine-rich peptide HSHRDFQPVLHL-NH(2) (L), identical with the N-terminal fragment of the anti-angiogenic human endostatin has been synthesized. Endostatin is a recently identified broad spectrum angiogenesis inhibitor, which inhibits 65 different tumor types. The N-terminal 25-mer peptide fragment of human endostatin has the same antitumor effect as the entire protein. The zinc(II) binding is crucial for the antitumor effect in both cases. Our peptide may provide all critical interactions for zinc(II) binding present in the N-terminal 25-mer peptide fragment. In addition, the N-terminus of human endostatin has a supposedly high affinity binding site for copper(II), similar to human serum albumin. Since copper(II) is intimately involved in angiogenesis, this may have biological relevance. In order to determine the metal binding properties of the N-terminal fragment of endostatin, we performed equilibrium, UV-visible (UV-vis), CD, EPR and NMR studies on the zinc(II) and copper(II) complexes of L. In the presence of zinc(II) the formation of a stable [NH(2),3N(im),COO(-)] coordinated complex was detected in the neutral pH-range. This coordination mode is probably identical to that present in the zinc(II) complex of the above mentioned N-terminal 25-mer peptide fragment of human endostatin. Moreover, L has extremely high copper(II) binding affinity, close to those of copper-containing metalloenzymes, and forms albumin-like [NH(2),N(-),N(-),N(im)] coordinated copper(II) complex in the neutral pH-range, which may suggest that copper(II) binding is involved in the biological activity of endostatin.
Kolozsi
András
A
Department of Inorganic and Analytical Chemistry, University of Szeged, Szeged, Hungary.
Jancsó
Attila
A
Nagy
Nóra V
NV
Gajda
Tamás
T
D020533:Angiogenesis Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D003300:Copper / Q000378:metabolism*; D043169:Endostatins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism*
D020533:Angiogenesis Inhibitors; D043169:Endostatins; D010446:Peptide Fragments; D003300:Copper; D015032:Zinc