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41.
Sizing large proteins and protein complexes by electrospray ionization mass spectrometry and ion mobility 总被引:1,自引:0,他引:1
Kaddis CS Lomeli SH Yin S Berhane B Apostol MI Kickhoefer VA Rome LH Loo JA 《Journal of the American Society for Mass Spectrometry》2007,18(7):1206-1216
Mass spectrometry (MS) and ion mobility with electrospray ionization (ESI) have the capability to measure and detect large noncovalent protein-ligand and protein-protein complexes. Using an ion mobility method of gas-phase electrophoretic mobility molecular analysis (GEMMA), protein particles representing a range of sizes can be separated by their electrophoretic mobility in air. Highly charged particles produced from a protein complex solution using electrospray can be manipulated to produce singly charged ions, which can be separated and quantified by their electrophoretic mobility. Results from ESI-GEMMA analysis from our laboratory and others were compared with other experimental and theoretically determined parameters, such as molecular mass and cryoelectron microscopy and X-ray crystal structure dimensions. There is a strong correlation between the electrophoretic mobility diameter determined from GEMMA analysis and the molecular mass for protein complexes up to 12 MDa, including the 93 kDa enolase dimer, the 480 kDa ferritin 24-mer complex, the 4.6 MDa cowpea chlorotic mottle virus (CCMV), and the 9 MDa MVP-vault assembly. ESI-GEMMA is used to differentiate a number of similarly sized vault complexes that are composed of different N-terminal protein tags on the MVP subunit. The average effective density of the proteins and protein complexes studied was 0.6 g/cm(3). Moreover, there is evidence that proteins and protein complexes collapse or become more compact in the gas phase in the absence of water. 相似文献
42.
The authors present a new potential energy curve, electric dipole moment function, and spin-orbit coupling function for OH in the X 2Pi state, based on high-level ab initio calculations. These properties, combined with a spectroscopically parametrized lambda-type doubling Hamiltonian, are used to compute the Einstein A coefficients and photoabsorption cross sections for the OH Meinel transitions. The authors investigate the effect of spin-orbit coupling on the lifetimes of rovibrationally excited states. Comparing their results with earlier ab initio calculations, they conclude that their dipole moment and potential energy curve give the best agreement with experimental data to date. The results are made available via EPAPS Document No. E-JCPSAG-017709. 相似文献
43.
This paper is concerned with a numerical solution of hyperbolic cooling tower shell, a class of full nonlinear problems in solid mechanics of considerable interest in engineering applications. In this analysis, the post-buckling analysis of cooling tower shell with discrete fixed support and under the action of wind loads and dead load is studied. The influences of ring-stiffener on instability load are also discussed. In addition, a new solution procedure for nonlinear problems which is the combination of load increment iteration with modified R-C are- length method is suggested. Finally, some conclusions having important significance for practice engineering are given.The Project Supported by National Natural Science Foundation of China. 相似文献
44.
Joseph A. Loo Peifeng Hu Patrick McConnell W. Tom Mueller Tomi K. Sawyer Venkataraman Thanabal 《Journal of the American Society for Mass Spectrometry》1997,8(3):234-243
The noncovalent binding of various peptide ligands to pp60src (Src) SH2 (Src homology 2) domain protein (12.9 ku) has been used as a model system for development of electrospray ionization mass spectrometry (ESI-MS) as a tool to study noncovalently bound complexes. SH2 motifs in proteins are critical in the signal transduction pathways of the tyrosine kinase growth factor receptors and recognize phosphotyrosine-containing proteins and peptides. ESI-MS with a magnetic sector instrument and array detection has been used to detect the protein-peptide complex with low-picomole sensitivity. The relative abundances of the multiply charged ions for the complex formed between Src SH2 protein and several nonphosphorylated and phosphorylated peptides have been compared. The mass spectrometry data correlate well to the measured binding constants derived from solution-based methods, indicating that the mass spectrometry-based method can be used to assess the affinity of such interactions. Solution-phase equilibrium constants may be determined by measuring the amount of bound and unbound species as a function of concentration for construction of a Scatchard graph. ESI-MS of a solution containing Src SH2 with a mixture of phosphopeptides showed the expected protein-phosphopeptide complex as the dominant species in the mass spectrum, demonstrating the method’s potential for screening mixtures from peptide libraries. 相似文献
45.
Modified calcium carbonate (CaCO3) nanoparticles with cubic- and spindle-like configuration were synthesized in situ by the typical bobbling (gas-liquid-solid) method. The modifiers, such as sodium stearate, octadecyl dihydrogen phosphate
(ODP) and oleic acid (OA), were used to obtain hydrophobic nanoparticles. The different modification effects of the modifiers
were investigated by measuring the active ratio, whiteness and the contact angle. Moreover, transmission electron microscopy
(TEM), X-ray diffraction (XRD) and thermogravimetry analysis (TGA analysis) were employed to characterize the obtained products.
A preliminary reaction mechanism was discussed. According to the results, the active ratio of CaCO3 modified by ODP was ca. 99.9% and the value of whiteness was 97.3% when the dosage of modifiers reached 2%. The contact angle
was 122.25° for the CaCO3 modified in the presence of sodium stearate, ODP and OA. When modified CaCO3 was filled into PVC, the mechanical properties of products were improved greatly such as rupture intensity, pull intensity
and fuse temperature. The compatibility and affinity between the modified CaCO3 nanoparticles and the organic matrixes were greatly improved.
Supported by the National Natural Science Foundation of China (Grant No. 50372025) 相似文献
46.
Shirley H. Lomeli Sheng Yin Rachel R. Ogorzalek Loo Joseph A. Loo 《Journal of the American Society for Mass Spectrometry》2009,20(4):593-596
Increased multiple charging of native proteins and noncovalent protein complexes is observed in electrospray ionization (ESI)
mass spectra obtained from nondenaturing protein solutions containing up to 1% (vol/vol) m-nitrobenzyl alcohol (m-NBA). The increases in charge ranged from 8% for the 690 kDa α7β7β7α7 20S proteasome complex to 48% additional charge for the zinc-bound 29 kDa carbonic anhydrase-II protein. No dissociation
of the noncovalently bound ligands/subunits was observed upon the addition of m-NBA. It is not clear if the enhanced charging is related to altered surface tension as proposed in the “supercharging” model
of Iavarone and Williams (Iavarone, A. T.; Williams, E. R. J. Am. Chem. Soc.
2003, 125, 2319–2327). However, more highly charged noncovalent protein complexes have utility in relaxing slightly the mass-to-charge
(m/z) requirements of the mass spectrometer for detection and will be effective for enhancing the efficiency for tandem mass spectrometry
studies of protein complexes. 相似文献
47.
48.
49.
Ogorzalek Loo RR Loo JA Du P Holler T 《Journal of the American Society for Mass Spectrometry》2002,13(7):804-812
Identities ascribed to the intact protein ions detected in MALDI-MS of whole bacterial cells or from other complex mixtures are often ambiguous. Isolation of candidate proteins can establish that they are of correct molecular mass and sufficiently abundant, but by itself is not definitive. An in vivo labeling strategy replacing methionine with selenomethionine has been employed to deliver an additional constraint for protein identification, i.e., number of methionine residues, derived from the shift in mass of labeled versus unlabeled proteins. By stressing a culture and simultaneously labeling, it was possible to specifically image the cells' response to the perturbation. Because labeled protein is only synthesized after application of the stress, it provides a means to view dynamic changes in the cellular proteome. These methods have been applied to identify a 15,879 Da protein ion from E. coli that was induced by an antibacterial agent with an unknown mechanism of action as SpY, a stress protein produced abundantly in spheroplasts. It has also allowed us to propose protein identities (and eliminate others from consideration) for many of the ions observed in MALDI (and ESI-MS) whole cell profiling at a specified growth condition. 相似文献
50.
利用两根极性不同的毛细柱,在全二维气相色谱上分析辽河油田遭受严重生物降解形成的稠油饱和烃组分,可以将传统色谱分析时形成的“基线鼓包”即不可分辨的复杂混合物(Unresolved Complex Mixtures)分开.根据饱和烃全二维气相色谱谱图的族分离特点和瓦片效应,结合飞行时间质谱提供的质谱信息初步解析不可分辨的复杂混合物主要成分.发现常规色谱分析时形成的所谓“基线鼓包”是由成千上万、含量相对较低的不同取代基的环状化合物组成,这些化合物在一维色谱上以分子量递增的顺序排列,在二维色谱上以极性的差异或者环的多少排列.C24之前的第一组不可分辨的复杂混合物主要由环己烷为基本单元的单环、双环和三环烷烃类化合物组成,信噪比在100以上的化合物数量约为饱和烃总数量的75%,质量分数是饱和烃总量的80%以上,是饱和烃的主要组成部分.C24之后出现的第二组不可分辨的复杂混合物主要由四个环或者五个环为基本单元的化合物组成,信噪比在100以上的化合物数量约为饱和烃总数量的17%,质量分数是饱和烃总量的0.5%.对稠油中这些不可分辨的复杂混合物的解析有助于对其成因机理的认识和高效开采方案的制定. 相似文献