本文選題：肌紅蛋白　切入點(diǎn)：人血清白蛋白　出處：《大連大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

【摘要】： 本論文首次用我們建立的熒光光譜法研究了光誘導(dǎo)下Mb(WT)和Mb(D44K)的去氧過程,比較了它們?cè)诠庹T導(dǎo)的去氧等方面的不同;又用紫外、熒光和圓二色光譜學(xué)方法研究了秋水仙堿與人血清白蛋白之間的相互作用。結(jié)果發(fā)現(xiàn): 1.熒光光譜法在研究光誘導(dǎo)的Mb去氧方面,比之拉曼、紫外等光譜法具有靈敏、方便、峰單一容易識(shí)別不被其它峰干擾等優(yōu)點(diǎn)。含氧Mb在光照下發(fā)生去氧作用,光照量越大,去氧量越大;光照下的溫度越高,去氧越快;N2可以把Mb活性中心鐵卟啉中的配位氧脫下來,因而可用熒光光譜法作含氧Mb與CO、CO2、H2、Cl2、NO、CS2等各種氣體相互作用的去氧研究。此外,熒光光譜法還可用作Mb中色氨酸和酪氨酸殘基與鐵卟啉之間的傳能研究。 2. Mb(D44K)的去氧速率比Mb(WT)慢,更加難于去氧。430nm激發(fā)時(shí),Mb(D44K)在597.9nm和628.8nm處出現(xiàn)兩個(gè)熒光發(fā)射峰,不同于Mb(WT)僅在597nm處出現(xiàn)一個(gè)熒光發(fā)射峰。經(jīng)研究證明628.8nm處熒光峰是Mb3+-H2O型中的H2O峰。光照也使此峰的熒光強(qiáng)度下降,但比去氧的速率慢。進(jìn)一步研究發(fā)現(xiàn),597nm處Mb(D44K)的熒光效率比Mb(WT)的熒光效率低。傳能實(shí)驗(yàn)表明Mb表面44位氨基酸由Asp突變?yōu)長(zhǎng)ys后,不影響Mb(D44K)中色氨酸和酪氨酸殘基傳遞給鐵卟啉的熒光效率,但使Mb(D44K)中色氨酸和酪氨酸殘基的熒光效率變高。430nm是研究Mb(WT)和Mb(D44K)光照去氧的最佳激發(fā)波長(zhǎng)。 3.秋水仙堿使人血清白蛋白的紫外吸收增強(qiáng),特征熒光峰猝滅,并且隨溫度升高猝滅常數(shù)Ksv降低。求算了不同溫度下秋水仙堿與人血清白蛋白相互作用的平衡常數(shù)與結(jié)合位點(diǎn)數(shù)。根據(jù)Van′t Hoff方程計(jì)算出?H = -11.66 KJ/mol,?S= 51.507J/mol·K,得出二者之間的作用力主要是靜電作用力。圓二色光譜測(cè)得加入秋水仙堿后,人血清白蛋白的α-螺旋降低,二級(jí)結(jié)構(gòu)改變。表明秋水仙堿對(duì)人血清白蛋白的熒光猝滅機(jī)制屬于形成配合物所引起的靜態(tài)猝滅。
[Abstract]:In this paper, we first studied the deoxygenation process of MbCWTand MbD44K) by using the fluorescence spectrometric method established by us for the first time, and compared their differences in photoinduced deoxygenation, and used UV, UV, UV, UV, UV, UV, UV, UV, UV, UV, UV, UV. The interaction between colchicine and human serum albumin was studied by fluorescence and circular dichroism. 1. The fluorescence spectrum method is more sensitive and convenient than Raman and ultraviolet spectroscopy in studying the deoxygenation of Mb induced by light, and it is easy to identify the single peak without interference by other peaks. The higher the amount of illumination is, the more the oxygen Mb is deoxidized under light. The higher the amount of deoxidization and the higher the temperature under light, the faster the deoxygenation can remove the coordination oxygen from the iron porphyrin in the active center of Mb. Therefore, the interaction of oxygen containing Mb with various gases, such as COO2CO2H2Cl2Cl2NOCS2 and so on, can be studied by fluorescence spectrometry. Fluorescence spectroscopy can also be used to study the energy transfer between tryptophan and tyrosine residues in Mb and iron porphyrin. 2. The deoxygenation rate of MbD44K) is slower than that of MbCWT), and it is more difficult to deoxidize D44K at 430nm. Two fluorescence emission peaks appear at 597.9 nm and 628.8 nm, respectively. The fluorescence peak at 628.8 nm is the H 2O peak in Mb3 H 2O type, and the fluorescence intensity of this peak is also decreased by illumination. But the rate of deoxygenation was slower than that of deoxygenation. The fluorescence efficiency of MbD44K at 597 nm was lower than that of MbCWT. the energy transfer experiment showed that 44 amino acids on Mb surface mutated from Asp to Lys. The fluorescence efficiency of tryptophan and tyrosine residues transferred to iron porphyrin was not affected, but the fluorescence efficiency of tryptophan and tyrosine residues in MbD44K) was increased by .430 nm. 3.Colchicine enhanced the ultraviolet absorption of human serum albumin and quenched the characteristic fluorescence peak. The equilibrium constants and binding sites of colchicine interacting with human serum albumin at different temperatures were calculated. The equilibrium constants and binding sites of colchicine and human serum albumin were calculated by Van't Hoff equation. H = -11.66 KJ / mol? S = 51.507 J / mol 路K, the main force between them was electrostatic force. The 偽 -helix of human serum albumin was decreased after colchicine was added to the circular dichroism spectrum. The change of secondary structure indicates that the fluorescence quenching mechanism of colchicine on human serum albumin belongs to the static quenching caused by the formation of complex.
【學(xué)位授予單位】：大連大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R341

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