Dy(III)(NR)3与鲱鱼精子DNA的相互作用研究：光谱与电化学分析

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本研究主要探讨了Dy(III)(NR)_3与鲱鱼精子DNA之间的相互作用，这是一种在光谱学和电化学手段下进行的深入探索。通过紫外-可见（UV-vis）光谱和荧光光谱的分析，研究人员揭示了Dy(III)(NR)_3与DNA之间的配位行为。双倒数法研究表明，两者之间的结合比例为1:1，这意味着一个Dy(III)(NR)_3分子与一个DNA分子形成稳定复合物。实验结果显示，在25℃时，Dy(III)(NR)_3与DNA的结合常数(Ka)为1.15×10^5 L/mol，而在35℃时，这个数值增加至2.09×10^5 L/mol。这些数值反映了随着温度升高，化合物与DNA的相互作用增强。热力学参数方面，反应的标准摩尔焓变（ΔrH_θ）为2.48×10^4 J/mol，表明这是一个放热过程；标准摩尔吉布斯自由能变（ΔrG_θ）为-2.34×10^4 J/mol，显示该反应在标准条件下是自发的；而标准摩尔熵变（ΔrS_θ）为161.7 J/(mol·K)，暗示着反应过程中有一定程度的熵增加。这项研究不仅提供了关于Dy(III)(NR)_3与鲱鱼精子DNA之间具体作用方式的信息，还对理解金属离子与生物大分子（如DNA）的相互作用机制具有重要意义。这类研究对于生物传感器、药物设计以及生物物理化学等领域具有潜在的应用价值。通过精细调控这种相互作用，可以为新型材料和生物技术的发展打开新的可能。

236 CHINESE OPTICS LETTERS / Vol. 8, No. 2 / February 10, 2010

Study on interaction between Dy(III)(NR)

complex

and herring sperm DNA by spectroscopy

Yan Zhang (ÜÜÜ ýýý)

, Xingming Wang (,,,²²²)

1∗

, Dan Fei (¤¤¤ ûûû)

, Na Zhao (ëëë AAA)

Tingting Zhao (ëëëxxxxxx)

, Haifeng Zhu (ÁÁÁ°°°¸¸¸)

, and Lisheng Ding (¶¶¶ááá))))

Department of Chemistry, Material Science and Engineering College,

Southwest University of Science and Technology, Mianyang 621010, China

Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China

∗

E-mail: xmwang xkd@yahoo.com.cn

Received March 23, 2009

The action mode between Dy(III)(NR)

and herring sperm DNA is studied by ultraviolet-visible (UV-

vis) and ﬂuorescence sp ectra as well as electrochemistry. Double-reciprocal method studies show that

the binding stoichiometry between Dy(III)(NR)

and DNA is 1 : 1, the binding constants at diﬀerent

temp eratures are 1.15 × 10

L/mol at 25

◦

C and 2.09 × 10

L/mol at 35

◦

C, and the corresponding

thermo dynamic parameters are ∆

= 2.48 × 10

J/mol, ∆

= −2.34 × 10

J/mol, ∆

= 161.7

J/(mol · K), ∆

is the driving force in this reaction. Combined with Scatchard method and melting

metho d, the results suggest that the interaction mode between Dy(III)(NR)

and herring sperm DNA is

intercalation fashion and groove fashion.

OCIS co des: 300.6390, 300.6280, 300.6550.

doi: 10.3788/COL20100802.0236.

DNA attracted much interest due to its crucial role in

gene expression, gene transcription, mutagenesis, car-

cinogenesis, and cell death, etc.

[1]

In recent years, stud-

ies on the interaction of small molecules with DNA have

gained prominence because of their relevance in the de-

velopment of new reagents for medicine. On one hand,

we can select new nucleic acid ﬂuorescence probe during

the study. On the other hand, some anticancer drugs

take cancer DNA as head target by ways of breaking its

structure and interfering its gene regulation and expres-

sion, showing anticancer activity

[2−5]

The noncovalent binding of some ligands may be fol-

lowed by chemical reaction or ligand-induced cleavage of

the nucleic acid. It is generally accepted that there are

three binding modes of small molecules to the DNA dou-

ble helix: intercalation binding, groove binding, and elec-

trostatic binding

[6−8]

. Intercalation, a strongly favorable

binding mode that can inﬂuence the function of DNA

by rupturing DNA directly or restraining its replication

and transcription, involves the π-stacking of a ligand be-

tween adjacent base pairs of DNA. This π-stacking in-

teraction requires the intercalating ligand to be a ﬂat,

extended aromatic compound, which is annulated with

heterocyclic rings, e.g., pyridine and pyrazine

[9]

. Neu-

tral red (NR), a planar phenazine dye, is sensitive to

the size, shape, chirality, and hydrophobic characteris-

tics of the complex. Compared with transition metals,

rare metals are more eﬀecive to cleave the phosphate di-

ester bond

[10−12]

. As a result, the study of their metal

complex Dy(III)(NR)

provides useful insights into drug

design and we can understand the binding mechanism of

these drugs.

Spectrophotometric measurements are known as pow-

erful tools for investigating the interaction between small

molecules and DNA. Although DNA has a natural ﬂu-

orescence, it is too weak to be used directly to mea-

sure DNA. Compared with a common ﬂuorimetric probe,

ethidium bromide (EB) and acridine orange (AO) oﬀer

lower toxicity, higher stability, and convenience of use.

In this letter, AO is selected as the probe

[13,14]

. The

interaction between Dy(III)(NR)

complex and herring

sperm DNA is conﬁrmed by spectrophotometer and elec-

trochemistry measurements. A series of thermo dynamic

parameters and binding constants are also obtained.

The complex was prepared by concentrating stoichio-

metric dysprosium chloride and NR in absolute ethanol,

recirculated on a water bath at 80

◦

C for 14 h. The

sample was dried in an oven for 5 h and remaining

the volume of about 10 mL. After standing for several

days, brownish crystals of Dy(III)(NR)

· H

[11]

were

yielded. Infrared (IR) spectra data display clearly the

symmetrical stretching band of NH at 3460 cm

−1

, the

NH bending band at 1625 cm

−1

, the stretching band

of CH in benzene ring at 860 cm

−1

, and the stretching

band of CN at 1170 and 1080 cm

−1

. Elemental analysis

shows the composition of C43.33 H4.71 N14.25 Dy14.38

in percentages

[15,16]

The herring sperm DNA (purchased from Sigma Bio-

logical Co.) was used without further puriﬁcation. The

DNA was dissolved in doubly distilled deionized water

with 50-mmol/L NaCl and dialyzed for 48 h against

a buﬀer solution at 4

◦

C. The purity of the DNA

was checked by monitoring the ratio of the 260-nm ab-

sorbance to 280-nm absorbance (A

260

280

). The ratio

of 1.83 indicated that DNA was fully free of protein

[17]

The DNA concentrations per nucleotide were determined

from the light absorbance at 260 nm by using the molar

extinction coeﬃcient of 6600 L/(mol·cm). The solution

of DNA was stored at 4

◦

C for a short time and then

used.

All of the samples were dissolved in tris-HCl buﬀer

(The concentration of tris was 0.1 mol/L by adding HCl

solution with the concentration of 0.1 mol/L. Acidometer

was used to examine the pH value of the buﬀer until it

1671-7694/2010/020236-05

° 2010 Chinese Optics Letters

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