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Positive selection is a general phenomenon in the evolution of abalone sperm lysin 总被引:36,自引:21,他引:15
Lysin is a 16kDa acrosomal protein used by abalone sperm to create a hole
in the egg vitelline envelope (VE). The interaction of lysin with the VE is
species-selective and is one step in the multistep fertilization process
that restricts heterospecific (cross-species) fertilization. For this
reason, the evolution of lysin could play a role in establishing prezygotic
reproductive isolation between species. Previously, we sequenced sperm
lysin cDNAs from seven California abalone species and showed that positive
Darwinian selection promotes their divergence. In this paper an additional
13 lysin sequences are presented representing species from Japan, Taiwan,
Australia, New Zealand, South Africa, and Europe. The total of 20 sequences
represents the most extensive analysis of a fertilization protein to date.
The phylogenetic analysis divides the sequences into two major clades, one
composed of species from the northern Pacific (California and Japan) and
the other composed of species from other parts of the world. Analysis of
nucleotide substitution demonstrates that positive selection is a general
process in the evolution of this fertilization protein. Analysis of
nucleotide and codon usage bias shows that neither parameter can account
for the robust data supporting positive selection. The selection pressure
responsible for the positive selection on lysin remains unknown.
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2.
Gene therapy has emerged as a promising strategy for treatment of various diseases, but there is a pressing need for the development of non-invasive reporter techniques based on appropriate molecules and imaging modalities to assay gene expression. We now report the design, synthesis, and evaluation of novel enhanced reporter molecules, which reveal lacZ gene expression: trifluoromethylated aryl beta-D-galactopyranosides. A series of five molecular structures were screened in solution and with stably transfected lacZ expressing human MCF7 breast cancer cells in vitro. p-Trifluoromethyl-o-nitrophenyl beta-D-galactopyranoside (PCF(3)ONPG) was found to exhibit valuable properties including a single (19)F NMR signal, stability in aqueous solution and with wild type cells, but a chemical shift response to enzyme cleavage (Deltadelta=1.14 ppm) in breast cancer cells transfected to stably express lacZ. 相似文献
3.
Mitochondrial DNA and bindin gene sequence evolution among allopatric species of the sea urchin genus Arbacia 总被引:3,自引:1,他引:2
Sea urchins of the genus Arbacia (order Stirodonta) have discontinuous
allopatric distributions ranging over thousands of kilometers.
Mitochondrial DNA (mtDNA) sequences were used to reconstruct phylogenetic
relationships of four Arbacia species and their geographic populations.
There is little evidence of genetic structuring of populations within
species, except in two cases at range extremes. The mtDNA sequence
differentiation between species suggests that divergence occurred about 4-9
MYA. Gene sequences encoding the sperm protein bindin and its intron were
obtained and compared with the mtDNA phylogeny. Sea urchins among the
well-studied echinoid order Camarodonta, with degrees of mtDNA divergence
similar to those of Arbacia species, are known to have remarkable variation
in bindin. However, in Arbacia, little variation in deduced amino acid
sequences of bindin was found, indicating that purifying selection acts on
the protein. In contrast, bindin intron sequences showed much
differentiation, including numerous insertion/deletions. Fertilization
experiments performed between a divergent pair of Arbacia species from the
Atlantic and Pacific Oceans revealed no evidence of blocks to gamete
recognition. In Arbacia, fertilization specificities may have evolved
relatively slowly as a result of extensive gene flow within species,
greater functional constraint on the bindin polypeptide, or reduced
selective pressure for species recognition in singly occurring species.
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4.
Females of the sea urchin Strongylocentrotus purpuratus differ in the structures of their egg jelly sulfated fucans 总被引:1,自引:0,他引:1
The egg jelly coats of sea urchins contain sulfated fucans which bind to a
sperm surface receptor glycoprotein to initiate the signal transduction
events resulting in the sperm acrosome reaction. The acrosome reaction is
an ion channel regulated exocytosis which is an obligatory event for sperm
binding to, and fusion with, the egg. Approximately 90% of individual
females of the sea urchin Strongylocentrotus purpuratus spawned eggs having
only one of two possible sulfated fucan electrophoretic isotypes, a slow
migrating (sulfated fucan I), or a fast migrating (sulfated fucan II)
isotype. The remaining 10% of females spawned eggs having both sulfated
fucan isotypes. The two sulfated fucan isotypes were purified from egg
jelly coats and their structures determined by NMR spectroscopy and
methylation analysis. Both sulfated fucans are linear polysaccharides
composed of 1-->3-linked alpha-L-fucopyranosyl units. Sulfated fucan I
is entirely sulfated at the O -2 position but with a heterogeneous
sulfation pattern at O -4 position. Sulfated fucan II is composed of a
regular repeating sequence of 3 residues, as follows: [3-alpha-L-Fuc p -
2,4(OSO3)-1-->3-alpha-L-Fuc p -4(OSO3)-1-->3-alpha-L-Fuc p -4(OSO3)-
1]n. Both purified sulfated fucans have approximately equal potency in
inducing the sperm acrosome reaction. The significance of two structurally
different sulfated fucans in the egg jelly coat of this species could
relate to the finding that the sperm receptor protein which binds sulfated
fucan contains two carbohydrate recognition modules of the C-type lectin
variety which differ by 50% in their primary structure.
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6.
Increased emphasis on personalized medicine and novel therapies requires the development of noninvasive strategies for assessing biochemistry in vivo. The detection of enzyme activity and gene expression in vivo is potentially important for the characterization of diseases and gene therapy. Magnetic resonance imaging (MRI) is a particularly promising tool, since it is noninvasive and has no associated radioactivity, yet penetrates deep tissue. We now demonstrate a novel class of dual (1)H/(19)F nuclear magnetic resonance (NMR) lacZ gene reporter molecule to specifically reveal enzyme activity in human tumor xenografts growing in mice. We report the design, synthesis, and characterization of six novel molecules and evaluation of the most effective reporter in mice in vivo. Substrates show a single (19)F NMR signal and exposure to β-galactosidase induces a large (19)F NMR chemical shift response. In the presence of ferric ions, the liberated aglycone generates intense proton MRI T(2) contrast. The dual modality approach allows both the detection of substrate and the imaging of product enhancing the confidence in enzyme detection. 相似文献
7.
Praveen K. Gulaka Federico Rojas-Quijano Zoltan Kovacs Ralph P. Mason A. Dean Sherry Vikram D. Kodibagkar 《Journal of biological inorganic chemistry》2014,19(2):271-279
Tumor hypoxia is known to affect sensitivity to radiotherapy and promote development of metastases; therefore, the ability to image tumor hypoxia in vivo could provide useful prognostic information and help tailor therapy. We previously demonstrated in vitro evidence for selective accumulation of a gadolinium tetraazacyclododecanetetraacetic acid monoamide conjugate of 2-nitroimidazole (GdDO3NI), a magnetic resonance imaging T 1-shortening agent, in hypoxic cells grown in tissue culture. We now report evidence for accumulation of GdDO3NI in hypoxic tumor tissue in vivo. Our data show that GdDO3NI accumulated significantly (p < 0.05) in the central, poorly perfused regions of rat prostate adenocarcinoma AT1 tumors (threefold higher concentration than for the control agent) and showed better clearance from well-perfused regions and complete clearance from the surrounding muscle tissue. Inductively coupled plasma mass spectroscopy confirmed that more GdDO3NI than control agent was retained in the central region and that more GdDO3NI was retained in the central region than at the periphery. These results show the utility of GdDO3NI to image tumor hypoxia and highlight the potential of GdDO3NI for application to image-guided interventions for radiation therapy or hypoxia-activated chemotherapy. 相似文献
8.
M. Meser Ali Mark Woods Eul Hyun Suh Zoltan Kovacs Gyula Tircsó Piyu Zhao Vikram D. Kodibagkar A. Dean Sherry 《Journal of biological inorganic chemistry》2007,12(6):855-865
Lanthanide complexes (Eu3+, Gd3+ and Yb3+) of two different 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tetraamide derivatives containing two (2) and four (3) O-benzyl-l-serine amide substituents were synthesized and their chemical exchange saturation transfer (CEST) and relaxometric properties
were examined in the presence and absence of human serum albumin (HSA). Both Eu2 and Eu3 display a significant CEST effect from a single slowly exchanging Eu3+-bound water molecule, making these PARACEST complexes potentially useful as vascular MRI agents. Yb2 also showed a detectable CEST effect from both the Yb3+-bound water protons and the exchangeable NH amide protons, making it potentially useful as a vascular pH sensor. Fluorescence
displacement studies using reporter molecules indicate that both Gd2 and Gd3 displace dansylsarcosine from site II of HSA with inhibition constants of 32 and 96 μM, respectively, but neither complex
significantly displaces warfarin from site I. Water proton relaxation enhancements of 135 and 171% were observed upon binding
of Gd2 and Gd3 to HSA, respectively, at 298 K and pH 7.4. 相似文献
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