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应用激光解吸质谱和十二烷基硫酸钠—聚丙烯酰胺凝胶电泳分析蛇毒?… 总被引:1,自引:0,他引:1
应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)和SDS-聚丙烯酰胺凝胶电流对吉林省两地市的同种白眉蝮蛇蛇毒中具有抗栓塞药效的精氨酸酯酶进行了分析了分析和比较。MALDI-TOF-MS法具有快速、准确度高、灵敏度高的优点,两种方法结合,互为补充,取得了令人满意的结果。MALDI-TOF-MS完全可以直接用作蛇毒成分分离过程中重要的研究手段。 相似文献
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采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)对杯芳烃化合物进行了分析。探讨了样品的制备条件、仪器操作参数等因素对测定结果的影响。25种样品的质谱数据表明,利用MALDI-TOF-MS可非常方便地得到灵敏度、分辨率、准确度均较高,且易于识别和解析的质谱图,为此类化合物的质谱表征提供和建立了一种新的高效分析方法。 相似文献
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采用MALDI-TOF-MS法快速测定了天花粉蛋白的分子量,并讨论和对比了三种不同基质对其影响,认为用基质芥子酸是最佳适宜条件。实验结果表明本方法优于其它传统的测定生物大分子分子量方法。 相似文献
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Perkins等[1]用MALDI/TOF/MS对不同种类蛇毒中神经毒素进行了分析,彭嘉柔等[2,3]对江浙蝮蛇和蛇毒粗组份进行了初步质谱表征.但蛇毒蛋白纯化困难,因此对单一组份的研究较少且不系统.本文以白眉蝮蛇蛇毒(AgkistrodonblomhoffiiUssurensis,ABUV)为原料,纯化得到了精氨酸酯酶(Arginineesterase,AEase)、磷脂酶A2(PhospholipaseA2,PLA2)、纤溶酶(fibrinolyticenzyme)和L-氨基酸氧化酶(L-aminoacidoxidase),并且用MALDI/TOF/MS法对它们和蛇毒粗毒进行了系统研究.1 实验部分1.1 仪器和药品 激光解吸质谱仪为美国Molecular公司L… 相似文献
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The lyophilized venom of the spider Agelenopsis aperta (Aranea: Agelenidae) has been re‐analyzed by on‐line coupled high‐performance liquid chromatography and atmospheric‐pressure chemical‐ionization mass spectrometry (HPLC‐UV(DAD)‐APCI‐MS and MS/MS). Thus, 33 acylpolyamines with 11 different molecular masses, present in various concentrations, were detected, and their structures were disclosed. Different types of possible fragmentation as well as principles of structure elucidation are discussed. The method can be used for rapid separation and analysis of spider‐venom constituents. 相似文献
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肿瘤的生长依赖于血管的生成,新生血管不仅为肿瘤生长提供必需的营养物质,而且为肿瘤细胞扩散提供了重要的途径。1997年哈佛大学的O'Reilly等发现了一种内源性新血管生成抑制因子内皮抑素(Endoscatin),显示出特异抑制激活的血管内皮细胞增殖和肿瘤新血管生成的生物学活性,其抗肿瘤作用具有高效、低毒、无耐药性的优点。目前,内皮抑素的研究引起了国内外广泛的兴趣,在美国已进行以安全性为目的的I期临床实验,国内也有多家公司对内皮抑素进行了抗肿瘤研究并申报一类新药。内皮抑素有望成为医治肿瘤而又没有化疗和放疗的毒副作用的一种新的治疗方法,但是否能作为药物应用于临床,尚需对内皮抑素的结构特点及抑制肿瘤和内皮细胞的作用机制等方面进行许多深入的研究。 相似文献
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Abdelwahab Khalil Basem H. Elesawy Tarek M. Ali Osama M. Ahmed 《Molecules (Basel, Switzerland)》2021,26(16)
Insects of the order Hymenoptera have a defensive substance that contains many biologically active compounds. Specifically, venom from honeybees (Apis mellifera) contains many enzymes and peptides that are effective against various diseases. Different research papers stated the possibility of using bee venom (a direct bee sting or in an injectable form) in treating several complications; either in vivo or in vitro. Other reports used the active fractions of bee venom clinically or at labratory scale. Many reports and publications have stated that bee venom and its constituents have multiple biological activities including anti-microbial, anti-protozoan, anti-cancer, anti-inflammatory, and anti-arthritic properties. The present review aims to refer to the use of bee venom itself or its fractions in treating several diseases and counteracting drug toxicities as an alternative protocol of therapy. The updated molecular mechanisms of actions of bee venom and its components are discussed in light of the previous updated publications. The review also summarizes the potential of venom loaded on nanoparticles as a drug delivery vehicle and its molecular mechanisms. Finally, the products of bee venom available in markets are also demonstrated. 相似文献
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John R. Perkins Brian Smith Richard T. Gallagher Davis S. Jones Stephen C. Davis Andrew D. Hoffman Kenneth B. Tomer 《Journal of the American Society for Mass Spectrometry》1993,4(8):670-684
Electrospray mass spectrometry (ES/MS) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF/MS) were used to provide mass spectra from seven elapid snake venoms. Spectral interpretation was much simpler for MALDI/TOF/MS. ES/MS proved more useful for the provision of molecular weight data for very closely related peptides, but suppression of higher molecular weight compounds was seen to occur during flow injection analysis. MALDI/TOF/MS proved useful for providing a complete picture of the venom, but the low resolution led to obscuring of major ions, and the mass accuracy was poorer for known peptides. Suppression also occurred during MALDI/TOF/MS but could be overcome using alternative matrices because the spectra were very dependent on the choice of matrix. ES/MS and MALDI/TOF/MS provide complementary and confirmatory information such that for the anal sis of complex peptide mixtures (snake venoms), the use of both techniques is desirable. 相似文献
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Quinton L Gilles N Smargiasso N Kiehne A De Pauw E 《Journal of the American Society for Mass Spectrometry》2011,22(11):1891-1897
This study describes the structural characterization of a totally new family of peptides from the venom of the snake green
mamba (Dendroaspis angusticeps). Interestingly, these peptides differ in several points from other already known mamba toxins. First of all, they exhibit
very small molecular masses, ranging from 1.3 to 2.4 kDa. The molecular mass of classical mamba toxins is in the range of
7 to 25 kDa. Second, the new peptides do not contain disulfide bonds, a post-translational modification commonly encountered
in animal toxins. The third difference is the very high proportion of proline residues in the sequence accounting for about
one-third of the sequence. Finally, these new peptides reveal a carbohydrate moiety, indicating a glycosylation in the sequence.
The last two features have made the structural characterization of the new peptides by mass spectrometry a real analytical
challenge. Peptides were characterized by a combined use of MALDI- TOF/TOF and nanoESI-IT-ETD experiments to determine not
only the peptide sequence but also the composition and the position of the carbohydrate moiety. Anyway, such small glycosylated
and proline-rich toxins are totally different from any other known snake peptide and form, as a consequence, a new family
of peptides. 相似文献
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K562细胞是一株分化差、恶性程度高的人白血病细胞。研究表明,在一些分化诱导剂的作用下,细胞可以向红细胞系、粒细胞系和巨K562核细胞系方向分化成熟,并表现出相应血细胞类型的成熟标志。用一维固相pH梯度等电聚焦电泳(IPGIEF)分离K562细胞总蛋白质,其每一条带常包含多个蛋白质,难以用肽指纹谱技术来鉴定。有人探索过基于液相色谱的串联质谱技术来鉴定混合蛋白质,但灵敏度上存在一些问题。本文尝试采用基质辅助激光解吸附电离-串联飞行时间质谱(MALDI-TOF-TOF)分析一维电泳条带,鉴定了K562细胞中部分可能与白血病相关的蛋白质。 相似文献
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Farid Jahouh Rina Saksena Donatella Aiello Anna Napoli Giovanni Sindona Pavol Kováč Joseph H. Banoub 《Journal of mass spectrometry : JMS》2010,45(10):1148-1159
We present the MALDI‐TOF/TOF‐MS analyses of various hapten–bovine serum albumin (BSA) neoglycoconjugates obtained by squaric acid chemistry coupling of the spacer‐equipped, terminal monosaccharide of the O‐specific polysaccharide of Vibrio cholerae O1, serotype Ogawa, to BSA. These analyses allowed not only to calculate the molecular masses of the hapten–BSA neoglycoconjugates with different hapten–BSA ratios (4.3, 6.6 and 13.2) but, more importantly, also to localize the covalent linkages (conjugation sites) between the hapten and the carrier protein. Determination of the site of glycation was based on comparison of the MALDI‐TOF/TOF‐MS analysis of the peptides resulting from the digestion of BSA with similar data resulting from the digestion of BSA glycoconjugates, followed by sequencing by MALDI‐TOF/TOF‐MS/MS of the glycated peptides. The product‐ion scans of the protonated molecules were carried out with a MALDI‐TOF/TOF‐MS/MS tandem mass spectrometer equipped with a high‐collision energy cell. The high‐energy collision‐induced dissociation (CID) spectra afforded product ions formed by fragmentation of the carbohydrate hapten and amino acid sequences conjugated with fragments of the carbohydrate hapten. We were able to identify three conjugation sites on lysine residues (Lys235, Lys437 and Lys455). It was shown that these lysine residues are very reactive and bind lysine specific reagents. We presume that these Lys residues belong to those that are considered to be sterically more accessible on the surface of the tridimensional structure. The identification of the y‐series product ions was very useful for the sequencing of various peptides. The series of a‐ and b‐product ions confirmed the sequence of the conjugated peptides. Copyright © 2010 John Wiley & Sons, Ltd. 相似文献
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金属离子对蛇毒蛋白生物活性及结构效应的影响 总被引:1,自引:0,他引:1
从旅顺产白眉蝮蛇(Gloydius blomhoffii brevicaudus, GBB) 蛇毒中纯化得到了3种电泳和质谱纯蛋白活性组分, 其酶解肽段采用高效液相色谱-电喷雾串联质谱(HPLC-nESI-MS/MS)进行序列测定, 与其它同源性蛇毒蛋白氨基酸序列比对发现, 3种蛋白为新的蛇毒磷脂酶A2、类凝血酶和金属蛋白酶, 分别将其命名为GBB-bPLA2, GBB-TLE和GBB-MP. 电感耦合等离子体发射光谱法(ICP-AES) 测得每个GBB-bPLA2和GBB-MP中含有一个Ca2+; 每个GBB-TLE中含有2个Zn2+. Ca2+可分别使GBB-bPLA2和GBB-MP的荧光发射波长向短波方向移动2.0和1.6 nm, 使二者的荧光发射强度提高14.0%和11.0%; Ca2+的存在可显著提高二者的热稳定性, 使GBB-bPLA2和GBB-MP的热变性温度分别提高1.5和2.0 ℃. Zn2+使GBB-TLE的荧光发射强度增高4.3%, 但对GBB-TLE的酯酶水解活性、荧光发射波长和热变性温度无显著影响. 金属离子的存在能够不同程度地影响蛇毒蛋白结构热稳定性, 但对蛇毒蛋白生物活性的作用则不同. 相似文献