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色度模型。含L-赖氨酸-L-缬氨酸的DNA与多肽之间的相互作用:圆二色性和热变性研究。

Chromatic models. Interactions between DNA and polypeptides containing L-lysine L-valine: circular dichroism and thermal denaturation studies.

作者信息

Mandel R, Fasman G D

出版信息

Biochemistry. 1976 Jul 13;15(14):3122-30. doi: 10.1021/bi00659a028.

Abstract

The interaction of calf thymus DNA with statistical copolymers of L-lysine and L-valine [poly(L-Lys100f-Lvalf)] and block copolymers [poly(L-Lys)100f-poly(L-Val)f] were investigated as a function of ionic strength using circular dichroism (CD) spectroscopy. It was found that valine suppresses the ability of the copolymer-DNA complexes to yield a psi-type CD spectra as found for poly(L-Lys)-DNA [Jordan, C.F., Lerman, L.S., and Venable, J.N. (1972), Nature (london), New Biol. 236, 67] and lowers the ionic strength at which CD distortion occurs. Thermal denaturation, simultaneously monitoring 280-nm ellipticity, [theta]280, and hyperchromicity, h280, was carried out on annealed complexes of poly(L-Lys)-DNA, poly(L-Lys84.5-L-Val15.5)-DNA, poly(L-Lys)87.2-poly(L-Val)12.8-DNA, and directly mixed complexes of poly(L-Lys)-DNA, IN 2.5 X 10(-4) MEDTA, pH 7.0 solution. The CD denaturation of uncomplexed DNA at several ionic strengths was also determined to examine pre-melting. Despite the inability of both statistical and block copolymers of L-Lys and L-Val to form psi-type complexes with DNA, they bind as well to DNA as does poly(L-Lys) and give rise to a thermal denaturation pattern showing bound peaks between 90 and 100 degrees C, seen clearly with CD denaturation. The thermal denaturation of mixed and annealed complexes of poly(L-Lys)-DNA shows similar patterns in hyperchromicity changes as a function of temperature but very different CD melts. From the CD melt of annealed poly(L-Lys)-DNA, it appears that aggregation and long-range order of the complex are significant in low salt (2.5 X 10(-4) MEDTA) as well as in 1.0 M NaCl. These studies further illustrate the importance of the nature of nonionic interactions (hydrophobic) between polypeptides and DNA in determining the behavior of their complexes, such as causing condensation into higher order asymmetric structures. In light of these observations, the possible significance to the CD melting of chromatin and the validity of identification of C-form DNA by CD spectroscopy are discussed.

摘要

利用圆二色光谱(CD)研究了小牛胸腺DNA与L-赖氨酸和L-缬氨酸的统计共聚物[聚(L-Lys100f-Lvalf)]以及嵌段共聚物[聚(L-Lys)100f-聚(L-Val)f]之间的相互作用与离子强度的关系。结果发现,缬氨酸抑制了共聚物-DNA复合物产生psi型CD光谱的能力,就像聚(L-Lys)-DNA那样[乔丹,C.F.,勒曼,L.S.,和维纳布尔,J.N.(1972),《自然》(伦敦),新生物学236,67],并降低了发生CD畸变的离子强度。对聚(L-Lys)-DNA、聚(L-Lys84.5-L-Val15.5)-DNA、聚(L-Lys)87.2-聚(L-Val)12.8-DNA的退火复合物以及聚(L-Lys)-DNA、IN 2.5×10(-4)MEDTA、pH 7.0溶液的直接混合复合物进行了热变性实验,同时监测280nm的椭圆率[θ]280和增色效应h280。还测定了几种离子强度下未复合DNA的CD变性,以研究预熔解情况。尽管L-Lys和L-Val的统计共聚物和嵌段共聚物都不能与DNA形成psi型复合物,但它们与DNA的结合能力与聚(L-Lys)相同,并产生热变性图谱,在90至100摄氏度之间出现结合峰,在CD变性中清晰可见。聚(L-Lys)-DNA的混合和退火复合物的热变性在增色效应随温度变化方面表现出相似的模式,但CD熔解非常不同。从退火的聚(L-Lys)-DNA的CD熔解情况来看,复合物的聚集和长程有序在低盐(2.5×10(-4)MEDTA)以及1.0M NaCl中都很显著。这些研究进一步说明了多肽与DNA之间非离子相互作用(疏水)的性质在决定其复合物行为(如导致凝聚成更高阶不对称结构)方面的重要性。根据这些观察结果,讨论了对染色质CD熔解的可能意义以及通过CD光谱鉴定C型DNA的有效性。

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