Comparative XANES study of serotransferrin and ovotransferrin at Cu K-edge: evidence of interactions among the metal sites

The Cu site structure of human serotransferrin and hen ovotransferrin using XANES spectroscopy has been investigated. Although the transferrin family proteins have been extensively studied, the results reported herein are the first concerning the structure of the metal site in C-terminal and N-terminal in the whole protein. Our structural data show that these proteins differ with regard to the independence of the two binding sites and the geometry of copper coordination, ranging from a poorly to a significantly distorted octahedron.     

Distribution and Cytogenetic Features of Triploid Males of Crucian Carp in Azov Basin

When studying uni-bisexual crucian carp (Carassius auratus gibelio) populations in the Azov basin in 1995–2000, we found triploid males, which constituted 2.5%, on average, of the total numbers of studied samples. The areas of nuclei of erythrocytes of triploid males were, on average, 1.35 times those in diploid males. At the same optical density of DNA, the sizes of mature spermatozoon heads in triploid males were, on average, 1.8 times smaller than in diploid males, as follows from the data obtained in summer 1996. The results of similar studies carried out during the period of natural spawning activity in 1997–1999 suggest that the sizes of spermatozoon heads in triploid males were, on the contrary, 1.5 those in diploid males. Triploid males were characterized by mosaicism of spermatozoon sizes and chromosome mosaicism in somatic cells. Electrophoretic analysis for the locus of transferrin confirmed the triploid status of this genetic group. The results of comparative crosses of crucian carps with different ploidy suggest a high fertilizing capacity of triploid males, as well as normal viability of their progenies. A distinct positive correlation (r = 0.73) was found between the numbers of triploid females and triploid males in mixed di-triploid populations. No significant correlation was found between males and females within di- and triploid forms.     

Age-Dependent Variation of Serum Protein Antigens in Sturgeons (Acipenseridae,Acipenseriformes)

We studied the variation of antigenic properties of proteins in two sturgeon species at different stages of postembryonic development. The deepest changes occurred in individual components of albumins and -globulins (transferrins) and were mostly related to an increased proportion of the protein accounting for these antigens. Transformation of the main component of albumins A1 into adult antigens was completed in 5-month old juveniles. The main component of -globulins A (component of transferrins) appeared in the blood flow much later than other proteins and could retain the juvenile features until the age of 3–4 years. Other antigens belonging to ₁- and ₂-globulins and the second component of transferrins were more stable and did not undergo substantial changes. The direction of ontogenetic variation of serum antigens in sturgeon juveniles did not depend on the habitat of adult fish in fresh or sea water.     

Aluminum site structure in serum transferrin and lactoferrin revealed by synchrotron radiation X-ray spectroscopy

The Al site structure of serum transferrin and lactoferrin is investigated using X-ray absorption near edge structure (XANES) spectroscopy. Al K-edge spectra in the mono- and dialuminum forms of the proteins have been recorded for the first time. Our results show that the aluminium ion is hexa-coordinated in an octahedral-like symmetry and that the monoaluminum form, where only the C-terminal binding site is saturated, has an increased structural distortion around the metal site.     

Egg-white and blood-serum proteins functioning by noncovalent interactions: Studies by chemical modification and comparative biochemistry

Some of the more interesting and important proteins are those that function by forming associations or complexes with other substances. The structure-function relationships of three of these with very different substances are transferrins, which chelate metal ions; avian ovomucoids, which form complexes with proteolytic enzymes; and antifreeze glycoproteins, which interact at the ice-solution interface. Interrelating studies on the comparative biochemistry with studies using chemical modification have helped identify the side-chain groups of the proteins involved in function as well as to be useful for studies on general protein chemistry. The most strongly associated interaction is the chelation of iron by transferrin, with an association constant of 10²¹; tyrosines, histidines, and sometimes aspartate are involved. For ovomucoids, individual substratelike residues such as lysine are involved in a Michaelis-like complex, and association constants are as high as 10¹⁰. By contrast, the antifreeze glycoproteins appear to function by a polymeric interaction at the surface of ice, wiht a much weaker association.This paper comes at the time of the senior author's 75th birthday and also at the time of 28 years of still ongoing research between both authors, who have worked in close association in their laboratory at the University of California, Davis since 1960.     

The toposome, essential for sea urchin cell adhesion and development, is a modified iron-less calcium-binding transferrin

We describe the structure and function of the toposome, a modified calcium-binding, iron-less transferrin, the first member of a new class of cell adhesion proteins. In addition to the amino acid sequence of the precursor, we determined by Edman degradation the N-terminal amino acid sequences of the mature hexameric glycoprotein present in the egg as well as that of its derived proteolytically modified fragments necessary for development beyond the blastula stage. The approximate C-termini of the fragments were determined by a combination of mass spectrometry and migration in reducing gels before and after deglycosylation. This new member of the transferrin family shows special features which explain its evolutionary adaptation to development and adhesive function in sea urchin embryos: (i) a protease-inhibiting WAP domain, (ii) a 280 amino acid cysteine-less insertion in the C-terminal lobe, and (iii) a 240 residue C-terminal extension with a modified cystine knot motif found in multisubunit external cell surface glycoproteins. Proteolytic removal of the N-terminal WAP domain generates the mature toposome present in the oocyte. The modified cystine knot motif stabilizes cell-bound trimers upon Ca-dependent dissociation of hexamer-linked cells. We determined the positions of the developmentally regulated cuts in the cysteine-less insertion, which produce the fragments observed previously. These fragments remain bound to the hexameric 22S particle in vivo and are released only after treatment of the purified toposome with reducing agents. In addition, some soluble smaller fragments with possible signal function are produced. Sequence comparison of five sea urchin species reveals the location of the cell-cell contact site targeted by the species-specific embryo dissociating antibodies. The evolutionary tree of 2-, 1-, and 0-ferric transferrins implies their evolution from a basic cation-activated allosteric design modified to serve multiple functions.     

Identification of Toxoplasma gondii proteins binding human lactoferrin: a new aspect of rhoptry proteins function

In this paper, we report on the isolation, purification and identification of two Toxoplasma gondii membrane proteins binding human lactoferrin. Parasite membrane proteins were isolated using the commercial Mem-PER Eukaryotic Membrane Protein Extraction System. After purification by lactoferrin affinity chromatography, three protein bands were detected with the molecular mass of 74, 63 and 58 kDa, two of which (63 and 58 kDa) specifically bound biotin labeled human lactoferrin as examined by competitive inhibition. Further identification of latter proteins by ESI/MS/MS amino acid sequencing technique revealed those proteins as Toxoplasma ROP4 (band 63 kDa) and ROP2 (band 58 kDa) antigens known to be involved in many mechanisms essential for the parasite pathogenicity, including host lactoferrin acquisition as determined in this study.     

Transferrin gene frequencies of feral and domestic goats in New Zealand

Abstract

Blood plasma from three populations of feral goats (Capra hircus L.) and from five domestic breeds was analysed by polyacrylamide gel electrophoresis. The albumin locus was monomorphic in all samples. The macroglobulin locus was variable but lacked resolution on the gel. The transferrin (Tf) locus was polymorphic. The only two Tf alleles observed (Tf A and Tf B) did not vary from those described by other authors. Gene frequencies of the two Tf alleles showed that one feral population could be distinguished from the other two. Gene frequencies of domestic breeds were variable and showed no definitive pattern. Domestic breed type could not be recognised in the feral populations on the basis of transferrin analysis alone.     

Immunological Comparison of Transferrins of Some European Cypirinid Fish

Double immunodiffusion in agar gel with antiserum to carp transferrin was used to prove immunological relatedness between the transferrins of some European cyprinid fish. Quantitative differences between transferrins of nine species of cyprinid fish were studied by micro-complement fixation with the use of entisera to transferrins of four species. Immunological distances were conspicuously great and amounted on average to about 125 immunological distance units (IDU). A high rate of evolution of transferrins wes obtained. namely 4.2 IDU/million years, which represents about 0.84% sequence differences per million years.