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1.
(1) Standardization of penicillinase has been made possible by the method for its assay. (2) A purified, dried and sterile penicillinase has been found to be a penicillin-inactivator superior to Clarase for the penicillin sterility test. (3) Preliminary studies show this penicillinase preparation may be used for inactivating penicillin in exudates of body fluids.  相似文献   

2.
A Wang  S D Lu  D F Mark 《Science (New York, N.Y.)》1984,224(4656):1431-1433
The gene encoding human interleukin-2 (IL-2) has been cloned from human spleen cells, peripheral blood lymphocytes, and the Jurkat cell line. Nucleotide sequence analysis of the gene revealed that the encoded IL-2 protein has three cysteines located at amino acid residues 58, 105, and 125 of the mature protein. Site-specific mutagenesis procedures were used to modify the IL-2 gene by changing each of the cysteine codons individually to serine codons. Substitution of serine for cysteine residues at either position 58 or 105 of the IL-2 protein substantially reduced biological activity, indicating that the cysteines at these positions are necessary for maintenance of the biologically active conformation and may therefore be linked by a disulfide bridge. The modified IL-2 protein containing a substitution at position 125 retained full biological activity, suggesting that the cysteine at this position is not involved in a disulfide bond and that a free sulfhydryl group at that position is not necessary for receptor binding.  相似文献   

3.
Absence of cystathionase in human fetal liver: is cystine essential?   总被引:4,自引:0,他引:4  
Cystathionase activity is not measurable in the livers of 24 human fetuses and 3 premature infants, and the concentration of cystathionine in the liver is higher than that of the brain. The placenta does not subserve the trans-sulfuration function. Cystine (or cysteine) thus may be an essential amino acid in the immature human.  相似文献   

4.
The v-sis oncogene encodes a platelet-derived growth factor (PDGF)-related product whose transforming activity is mediated by its functional interaction with the PDGF receptor. PDGF, as well as processed forms of the v-sis gene product, is a disulfide-linked dimer with eight conserved cysteine residues in the minimum region necessary for biologic activity. Site-directed mutagenesis of the v-sis gene revealed that each conserved cysteine residue was required directly or indirectly for disulfide-linked dimer formation. However, substitution of serine for cysteine codons at any of four positions had no detrimental effect on transforming activity of the encoded v-sis protein. These results establish that interchain disulfide bonds are not essential in order for this protein to act as a functional ligand for the PDGF receptor. The remaining four substitutions of serine for cysteine each inactivated transforming function of the molecule. In each case this was associated with loss of a conformation shown to involve intramolecular disulfide bonds. These studies provide insight into the role of individual cysteine residues in determining the structure of the sis/PDGF molecule critical for biological activity.  相似文献   

5.
Several methanogenic archaea lack cysteinyl-transfer RNA (tRNA) synthetase (CysRS), the essential enzyme that provides Cys-tRNA(Cys) for translation in most organisms. Partial purification of the corresponding activity from Methanocaldococcus jannaschii indicated that tRNA(Cys) becomes acylated with O-phosphoserine (Sep) but not with cysteine. Further analyses identified a class II-type O-phosphoseryl-tRNA synthetase (SepRS) and Sep-tRNA:Cys-tRNA synthase (SepCysS). SepRS specifically forms Sep-tRNA(Cys), which is then converted to Cys-tRNA(Cys) by SepCysS. Comparative genomic analyses suggest that this pathway, encoded in all organisms lacking CysRS, can also act as the sole route for cysteine biosynthesis. This was proven for Methanococcus maripaludis, where deletion of the SepRS-encoding gene resulted in cysteine auxotrophy. As the conversions of Sep-tRNA to Cys-tRNA or to selenocysteinyl-tRNA are chemically analogous, the catalytic activity of SepCysS provides a means by which both cysteine and selenocysteine may have originally been added to the genetic code.  相似文献   

6.
The 21-kD proteins encoded by ras oncogenes (p21Ras) are modified covalently by a palmitate attached to a cysteine residue near the carboxyl terminus. Changing cysteine at position 186 to serine in oncogenic forms produces a nonpalmitylated protein that fails to associate with membranes and does not transform NIH 3T3 cells. Nonpalmitylated p21Ras derivatives were constructed that contained myristic acid at their amino termini to determine if a different form of lipid modification could restore either membrane association or transforming activity. An activated p21Ras, altered in this way, exhibited both efficient membrane association and full transforming activity. Surprisingly, myristylated forms of normal cellular Ras were also transforming. This demonstrates that Ras must bind to membranes in order to transmit a signal for transformation, but that either myristate or palmitate can perform this role. However, the normal function of cellular Ras is diverted to transformation by myristate and therefore must be regulated ordinarily by some unique property of palmitate that myristate does not mimic. Myristylation thus represents a novel mechanism by which Ras can become transforming.  相似文献   

7.
邻二氮菲-铁(Ⅲ)测定半胱氨酸   总被引:1,自引:0,他引:1  
研究了邻二氮菲-Fe3+与半胱氮酸的反应,发现半胱氨酸浓度在1.0×10-5~1.8×10-4mol.L-1范围内其产物吸光度值符合朗伯比耳定律。检测限1.0×10-6mol.L-1,表观摩尔吸光系数7.9×103L.mol-1.cm-1。采用本法测定谷氨酸、赖氨酸、半胱氨酸混合试样中半胱氨酸含量,相对标准偏差≤2%,结果令人满意。  相似文献   

8.
Thioglycolic acid neutralized with sodium carbonate, sodium thioglycolate (Eastman), ethyl mercaptan, cysteine hydrochloride and certain sulfur-containing reducing agents (sodium bisulfite and sodium hydrosulfite) antagonize the antibacterial action of 2-methyl-1,4-naphthoquinone on Escherichia coli in a synthetic medium. Other reducing agents such as stannous chloride, potassium formate and sodium thiosulfate, show no such antagonism. The antibacterial activities of 2-methyl-3-chloro-1,4-naphthoquinone and 2,6-dimethyl-1,4-naphthoquinone are also abolished by excess thioglycolate and cysteine, while that of 2-methyl-3-methoxy-1,4-naphthoquinone with -OCH(3) instead of -Cl or -H in the 3 position on the quinone ring, is not. These findings suggest that the mode of antibacterial action of 2-methyl-1,4-naphthoquinone is by blocking essential enzymes through combination with sulfhydryl groups, or through combination with sulfhydryl groups of essential bacterial metabolites. This combination may take place in the 3-position on the quinone ring. This mode of action is similar to that suggested by other investigators for several antibiotic agents including penicillin. The antibacterial activity of the methoxy quinone, however, even in the presence of sulfhydryl groups, suggests that the foregoing explanation may not be the complete one.  相似文献   

9.
Oxidation of D-glucose-C(14) by bovine thyroid slices was stimulated by cysteine, 2-aminoethanethiol, and cystamine. Amino acids tested, other than cysteine, were not stimulatory, except tyrosine and arginine to a slight degree. The pattern of stimulation resembled that brought about by thyroid-stimulating hormone. Stimulation of labeled CO(2) production was greater when glucose-1-C(14) rather than glucose-6-C(14) was the substrate.  相似文献   

10.
LON GW  FARAH A 《Science (New York, N.Y.)》1946,104(2697):220-221
The acute toxicity of an organic mercurial as studied on the mouse, intact anesthetized cat and dog, and the heart-lung preparation of the dog can be readily counteracted by sulfhydryl-containing compounds such as glutathione, cysteine, and 2,3-dimercapto-propanol. Equimolar doses of 2,3-dimercapto-propanol are about five times as active as cysteine or glutathione.  相似文献   

11.
Directed mutagenesis of dihydrofolate reductase   总被引:17,自引:0,他引:17  
Three mutations of the enzyme dihydrofolate reductase were constructed by oligonucleotide-directed mutagenesis of the cloned Escherichia coli gene. The mutations--at residue 27, aspartic acid replaced with asparagine; at residue 39, proline replaced with cysteine; and at residue 95, glycine replaced with alanine--were designed to answer questions about the relations between molecular structure and function that were raised by the x-ray crystal structures. Properties of the mutant proteins show that Asp-27 is important for catalysis and that perturbation of the local structure at a conserved cis peptide bond following Gly-95 abolishes activity. Substitution of cysteine for proline at residue 39 results in the appearance of new forms of the enzyme that correspond to various oxidation states of the cysteine. One of these forms probably represents a species cross-linked by an intrachain disulfide bridge between the cysteine at position 85 and the new cysteine at position 39.  相似文献   

12.
本文研究了半胱氨酸在室温下与磷钼杂多酸的反应,产物在P~H=9.8的氨一氯化铵缓冲溶液中有一灵敏的导数阳极吸附波可用于半胱氨酸的定量测定。半胱氨酸浓度在1×10~8mol/L~5×10~(-5)mol/L与导数波高有线性关系,检测限为5×10~(-9)mol/L。对混合样品中半胱氨酸的测定,回收率在94.5%~99.3%之间变化,相对标准偏差分别为0.4%和0.3%  相似文献   

13.
The active-site cysteine of peroxiredoxins is selectively oxidized to cysteine sulfinic acid during catalysis, which leads to inactivation of peroxidase activity. This oxidation was thought to be irreversible. However, by metabolic labeling of mammalian cells with 35S, we show that the sulfinic form of peroxiredoxin I, produced during the exposure of cells to H2O2, is rapidly reduced to the catalytically active thiol form. The mammalian cells' ability to reduce protein sulfinic acid might serve as a mechanism to repair oxidatively damaged proteins or represent a new type of cyclic modification by which the function of various proteins is regulated.  相似文献   

14.
[目的]对无核荔枝的半胱氨酸蛋白酶抑制剂基因进行克隆,并对其序列下进行分析。[方法]根据构建的无核荔枝胚败育SSH消减文库的半胱氨酸蛋白酶抑制剂EST序列,通过RACE技术获得半胱氨酸蛋白酶抑制基因的核苷酸序列并应用生物信息学软件进行分析。[结果]获得一个635 bp的半胱氨酸蛋白酶抑制基因序列,预测该序列含有321 bp的开放阅读框,推导其编码的蛋白质含106个氨基酸,具有半胱氨酸蛋白酶抑制剂保守区,与多个物种的半胱氨酸蛋白酶抑制剂基因具有较高的同源性。[结论]为进一步研究半胱氨酸蛋白酶抑制剂在植物中的生理功能奠定了基础。  相似文献   

15.
金属硫蛋白(Metallothionein简称MT)可通过半胱氨酸残基的巯基与金属离子结合形成无毒或低毒的络合物,从而消除重金属的毒害作用。为研究绊根草二型金属硫蛋白CdMTII络合AsO33-的能力强弱及其具有的特性,把目的基因转化到模式生物酵母中,通过酵母转化体表现的抵抗重金属的能力来检测目的基因的功能强弱,为该基因转化到植物体中培育可用于植物修复的工程植物品种以及在医学等其它领域的应用奠定基础。  相似文献   

16.
植物半胱氨酸合成酶复合体(SAT/OAS-TL)研究进展   总被引:1,自引:0,他引:1  
综述了植物半胱氨酸的合成调节和半胱氨酸合成酶复合体的结构、调节机理及其植物转基因的研究进展。  相似文献   

17.
The dipeptide sequence at the carboxy-terminal of a heavy (micro) chain from a human macroglobulin ( IgM) is tyrosylcysteine, although the reverse sequence, cysteinyltyrosine, has not been rigorously excluded. The presence of cysteine at the carboxy-terminal was predicted from a recognition of the chemical homologies among the polypeptide chains of immunoglobulins, and their probable evolutionary origin.  相似文献   

18.
Cell surface molecules of eukaryotic cells have been considered to be integrated into the membrane bilayer by a transmembrane protein sequence. The Thy-1 antigen of rodent thymocytes and brain was the first eukaryotic membrane molecule for which biochemical data clearly suggested membrane integration via a nonprotein tail. Direct evidence is now presented showing that a glycophospholipid structure is attached to the carboxyl-terminal cysteine residue and that 31 carboxyl-terminal amino acids predicted from the Thy-1 complementary DNA sequence are not present in the mature glycoprotein. These experimental results raise questions concerning signaling across a cell membrane since antibodies to Thy-1 can stimulate T lymphocytes to release lymphokines and undergo cell division.  相似文献   

19.
本研究通过固相合成法合成芋螺毒素Lv68线性肽,研究并优化了其主要异构体的一步氧化折叠条件,并通过电压膜片钳技术对主异构体产物进行了多种乙酰胆碱受体亚型(α3β4,α6β4,α4β2,α9α10,α1β1γδ,α1β1δε nAChRs)的活性测试,旨在为以烟碱型乙酰胆碱受体为靶点的先导药物分子的发现奠定基础,同时为该类其他芋螺毒素的氧化折叠方法、生物活性研究以及深入开发利用提供参考和借鉴。结果表明,芋螺毒素Lv68的一步法氧化折叠最优条件为反应体系组成为0.1 mol·L?1 Tris-HCl,1 mmol·L?1 EDTA,GSH/GSSG 1 mmol·L?1/1 mmol·L?1,线性肽浓度为50 μmol·L?1,反应时间为2 h,产率高达18.14 %,Lv68折叠肽在1 μmol·L?1浓度下对α9α10 nAChR具有较好的阻断作用(阻断率约80%)。本研究明确了芋螺毒素Lv68的一步氧化折叠条件,证实了Lv68对α9α10乙酰胆碱受体亚型具有阻断活性,拓宽了含半胱氨酸框架III的M超家族芋螺毒素的作用靶点范围。  相似文献   

20.
Parathyroid hormone loses biological activity upon oxidation with hydrogen peroxide. Part or all of this lost activity can be regained by subsequent reduction with cysteine. The extent and reversibility of this oxidation is dependent upon pH.  相似文献   

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