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51.
Heterogeneity of the glutathione transferase genes encoding enzymes responsible for insecticide degradation in the housefly 总被引:6,自引:0,他引:6
Michael Syvanen Zonghan Zhou Jonathan Wharton Claire Goldsbury Alan Clark 《Journal of molecular evolution》1996,43(3):236-240
One of the four glutathione-S-transferases (GST) that is overproduced in the insecticide-resistant Cornell-R strain of the housefly (Musca domestica) produces an activity that degrades the insecticide dimethyl parathion and conjugates glutathione to lindane. In earlier
work, it was shown that the resistant Cornell-R carries an amplification, probably a duplication, of one or more of its GST
loci and that this amplification is directly related to resistance. Using polymerase chain reaction (PCR) amplification with
genomic DNA, multiple copies of the gene encoding the parathion-degrading activity (called MdGst-3) were subcloned from both
the ancestral, insecticide-susceptible strain BPM and from the insecticide-resistant Cornell-R. In BPM, three different MdGst-3
genes were identified while in Cornell-R, 12 different MdGst-3 sequences were found that, though closely related to ancestral
genes, had diverged by a few nucleotides. This diversity in MdGst-3 genomic sequences in Cornell-R is reflected in the expressed
sequences, as sampled through a cDNA bank. Population heterozygosity cannot account for these multiple GST genes. We suggest
that selection for resistance to insecticides has resulted in not only amplification of the MdGst-3 genes but also in the
divergence of sequence between the amplified copies.
Received: 22 November 1995 / Accepted: 23 February 1996 相似文献
52.
The small (116 amino acids) inner membrane protein MerT encoded by the transposon Tn501 has been overexpressed under the control of the bacteriophage T7 expression system. Random mutants of MerT were made and screened for loss of mercuric ion hypersensitivity. Several mutantmerT genes were selected and sequenced: Cys24Arg and Cys25Tyr mutations abolish mercury resistance, as do charge-substitution mutations in the first predicted transmembrane helix (Glyl4Arg, Glyl5Arg, Gly27Arg, Ala18Asp), and the termination mutations Trp66Ter and Cys82Ter. 相似文献
53.
Jonathan A. Lindquist Elisabeth Barofsky Philip N. McFadden 《Journal of Protein Chemistry》1996,15(1):115-122
Protein (d-aspartyl/l-isoaspartyl) carboxyl methyltransferase (PCM, E.C. 2.1.1.77) was previously shown to be enzymatically methyl esterified in an autocatalytic manner at altered aspartyl residues; methyl esters are observed in a subpopulation of the enzyme termed thePCM fraction [Lindquist and McFadden (1994),J. Protein Chem.
13, 23–30]. The altered aspartyl sites serving as methyl acceptors inPCM have now been localized by using proteolytic enzymes and chemical cleavage techniques in combination with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry to identify fragments of the [3H]automethylated enzyme that contain a [3H]methyl ester. Methylation was positively identified at positions Asn188 and Asp217 in the enzyme sequence, a consequence of the spontaneous alteration of these sites tol-isoaspartyl ord-aspartyl sites and their methylation by active PCM molecules. The identification of more than one site of automethylation shows thatPCM is not a homogeneous population of damaged PCM molecules, but rather a complex population of molecules with a variety of age-altered damage sites.Abbreviations PCM
protein (d-aspartyl/l-isoaspartyl) carboxyl methyltransferase
- EDTA
disodium ethylenediaminetetraacetate
- PMSF
phenylmethylsulfonyl fluoride
- TEA
trifluoroacetic acid
- HPLC
high-pressure liquid chromatography 相似文献
54.
Sojourners: The Return of German Jews and the Question of Identity. John Borneman and Jeffrey M. Peck. Lincoln
Recovered Roots: Collective Memory and the Making of Israeli National Tradition. Yael Zerubavel. Chicago
The Masada Myth: Collective Memory and Mythmaking in Israel. Nachman Ben-Yehuda. Madison 相似文献
Recovered Roots: Collective Memory and the Making of Israeli National Tradition. Yael Zerubavel. Chicago
The Masada Myth: Collective Memory and Mythmaking in Israel. Nachman Ben-Yehuda. Madison 相似文献
55.
56.
A question that is central to understanding the mechanisms of aging and cellular deterioration is whether enzymes involved in recognition and metabolism of spontaneously damaged proteins are themselves damaged, either becoming substrates for their own activity; or being unable to act upon themselves, initiating cascades of cellular damage. We show here byin vitro experiments that protein (d-aspartyl/l-isoaspartyl) carboxyl methyltransferase (PCM) from bovine erythrocytes does methylate age-dependent amino acid damage in its own sequence. The subpopulation that is methylated, termed thePCM fraction, appears to be formed through age-dependent deamidation of an asparaginyl site to either anl-isoaspartyl ord-aspartyl site because (a) the stoichiometry of automethylation of purified PCM is less than 1%, a value typical of the substoichiometric methylation of many other aged protein substrates, (b)PCM is slightly more acidic than the bulk of PCM, and (c) the methyl esterified site inPCM has the characteristic base-lability of this type of methyl ester. Also, the methyl group is not incorporated into the enzyme as an active site intermediate because the incorporated methyl group is not chased onto substrate protein. The effect of enzyme dilution on the rate of the automethylation reaction is consistent with methylation occurring between protein molecules, showing that the pool of PCM is autocatalytic even though individual molecules may not be. The automethylation and possible self-repair of the PCM pool has implications for maintaining thein vivo efficiency of methylation-dependent protein repair. 相似文献
57.
58.
Catherine L. Craig Gary D. Bernard Jonathan A. Coddington 《Evolution; international journal of organic evolution》1994,48(2):287-296
We measured the reflectance properties of unpigmented silks spun by a systematic array of primitive (Deinopoidea) and derived (Araneoidea) aerial, web-spinning spiders, as well as silks spun by Araneomorphae and Mygalomorphae spiders that do not spin aerial webs. Our data show that all of the primitive aerial web spinners produce catching silks with a spectral peak in the ultraviolet (UV), and cladistic analysis suggests that high UV reflection is the primitive character state for silk spectral properties. In contrast, all of the derived aerial web spinners produce silks that are spectrally flat or characterized by reduced reflectance in the UV. Correlated with the evolution of these catching silks is a 37-fold increase in species number and apparent habitat expansion. This suggests that the unique silk proteins spun by the araneoids have been important to their ecological and evolutionary diversity. 相似文献
59.
Jonathan A. Sherratt 《Bulletin of mathematical biology》1994,56(1):129-146
More than 20 years after its proposal, Keller and Segel's model (1971,J. theor. Biol.,30, 235–248) remains by far the most popular model for chemical control of cell movement. However, before the Keller-Segel equations
can be applied to a particular system, appropriate functional forms must be specified for the dependence on chemical concentration
of the cell transport coefficients and the chemical degradation rate. In the vast majority of applications, these functional
forms have been chosen using simple intuitive criteria. We focus on the particular case of eukaryotic cell movement, and derive
an approximation to the detailed model of Sherrattet al. (1993,J. theor. Biol.,162, 23–40). The approximation consists of the Keller-Segel equations, with specific forms predicted for the cell transport coefficients
and chemical degradation rate. Moreover, the parameter values in these functional forms can be directly measured experimentally.
In the case of the much studied neutrophil-peptide system, we test our approximation using both the Boyden chamber and under-agarose
assays. Finally, we show that for other cell-chemical interactions, a simple comparison of time scales provides a rapid check
on the validity of our Keller-Segel approximation. 相似文献
60.
The functional units of a peptostreptococcal protein L 总被引:4,自引:0,他引:4
Jonathan P. Murphy Clive J. Duggleby Max A. Atkinson † Angus R. Trowern Tony Atkinson Christopher R. Goward 《Molecular microbiology》1994,12(6):911-920
Protein L is a cell-surface protein from Peptostreptococcus which interacts with immunoglobulin kappa light chains. A gene from Peptostreptococcus strain 3316 coding for protein L and fragments thereof were expressed in Escherichia coli. The peptides were examined for binding to immunoglobulin and serum albumin. The four C units were shown to be responsible for binding to immunoglobulin and the four D units for binding to albumin. This protein L molecule therefore binds to albumin at a site separate from that involved in binding to immunoglobulin. The albumin-binding units have high amino acid sequence identity with the albumin-binding units of streptococcal cell-surface proteins. The gene contains three sites available for internal initiation of translation resulting in three active proteins. The protein L molecule presented in this report was compared with a previously reported protein from Peptostreptococcus strain 312. The two proteins differ in several respects, including size and the number and types of repeat units. 相似文献