title: Human alpha-fetoprotein as a Zn(2+)-binding protein. Tight cation binding is not accompanied by global changes in protein structure and stability.
abstract: The binding of zinc to human alpha-fetoprotein (AFP) isolated from human umbilical cord serum was studied by fluorimetric Zn(2+)-titration. We found that the total number of strong binding sites for zinc on this protein was 5: AFP has one very strong (dissociation constant, K(d)<10(-8) M) and at least four lower affinity zinc binding sites (K(d)<10(-5) M). Fourier transform infrared (FTIR) analysis revealed that aspartate and histidine residues could be involved in the strong coordination of zinc. Intriguingly, binding of zinc to the protein does not induce structural changes that can be detected by circular dichroism, FTIR, intrinsic fluorescence or (1,1')-bi-(4-anilino)naphthalene-5,5'-disulfonic acid (bis-ANS) binding. Finally, scanning microcalorimetry measurements showed that stability of the protein is also unaffected by zinc binding in spite of the strength of the coordination. Such strong interactions without major structural consequences are highly unusual, and AFP may therefore be the first characterized representative of a new class of ligand-binding proteins.
authors.affiliation: Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino, Moscow Region, Russia.
authors.forename: Serge E
authors.initials: SE
authors.lastname: Permyakov
authors.forename: Keith A
authors.initials: KA
authors.lastname: Oberg
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: Cherskaya
authors.forename: Mikhail M
authors.initials: MM
authors.lastname: Shavlovsky
authors.forename: Eugene A
authors.initials: EA
authors.lastname: Permyakov
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Uversky
mesh_terms: D002152:Calorimetry, Differential Scanning; D002352:Carrier Proteins / Q000737:chemistry*; D002413:Cations, Divalent; D002942:Circular Dichroism; D005312:Fetal Blood; D005470:Fluorometry; D006801:Humans; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared; D015032:Zinc / Q000737:chemistry*; D000509:alpha-Fetoproteins / Q000737:chemistry*
chemical_list: D002352:Carrier Proteins; D002413:Cations, Divalent; D000509:alpha-Fetoproteins; C017800:zinc-binding protein; D015032:Zinc