title: X-ray absorption fine structure as a monitor of zinc coordination sites during oogenesis of Xenopus laevis.
abstract: The x-ray absorption fine structure (XAFS) zinc K-edge steps for intact stages I,II and V,VI Xenopus laevis oocytes demonstrate that the zinc concentration is about 3 and 1 mM, respectively. However, the chi(k) function for the early stage oocytes differs markedly from that for the late one. Analysis of the XAFS data for stage I,II oocytes indicates that zinc is bound to 2.0 +/- 0.5 sulfur atoms at an average coordination distance of 2.29 +/- 0.02 angstroms and 2.0 +/- 0.5 nitrogen or oxygen (N/O) atoms at 2.02 +/- 0.02 angstroms. In marked contrast, in stage V,VI oocytes, zinc is bound to 4.1 +/- 0.4 N/O atoms at an average distance of 1.98 +/- 0.01 angstroms. Our previous studies demonstrated that 90% of the zinc in stage VI oocytes is sequestered within yolk platelets, associated with a single molecule, lipovitellin, the proteolytically processed product of vitellogenin. XAFS analysis of yolk platelets, lipovitellin, and vitellogenin demonstrates that zinc is bound to 4.0 +/- 0.5 N/O ligands at an average distance of 1.98 +/- 0.01 angstroms in each case, identical to that of stage V,VI oocytes. The higher shell contributions in the Fourier transforms indicate that two of the N/O zinc ligands are His in both stage V,VI and I,II oocytes. The results show that in stage I,II oocytes, there is a high concentration of a zinc protein whose zinc coordination site likely is composed of (His)2(Cys)2, such as, e.g., TFIIIA. As the oocytes develop, the predominant zinc species becomes one that exhibits the (His)2(N/0)2 zinc site found in lipovitellin. Hence, the ligands to the zinc atoms in intact oocytes and the changes that take place as a function of oogenesis and after their fertilization, during embryogenesis, now can be examined and explored.
authors.affiliation: Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: D S
authors.initials: DS
authors.lastname: Auld
authors.forename: K H
authors.initials: KH
authors.lastname: Falchuk
authors.forename: K
authors.initials: K
authors.lastname: Zhang
authors.forename: M
authors.initials: M
authors.lastname: Montorzi
authors.forename: B L
authors.initials: BL
authors.lastname: Vallee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004527:Egg Proteins; D015387:Egg Proteins, Dietary / Q000235:genetics / Q000378:metabolism; D005260:Female; D066298:In Vitro Techniques; D008024:Ligands; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism*; D009866:Oogenesis / Q000502:physiology*; D013052:Spectrometry, X-Ray Emission; D014819:Vitellogenins / Q000235:genetics / Q000378:metabolism; D014982:Xenopus laevis; D015032:Zinc / Q000378:metabolism*
chemical_list: D004527:Egg Proteins; D015387:Egg Proteins, Dietary; D008024:Ligands; D014819:Vitellogenins; C001549:lipovitellin; D008666:Metalloendopeptidases; D015032:Zinc