Application of chromatographic retention data in an investigation of a quantitative structure-nucleotide incorporation rate relationship

K. Valkó, T. Cserháti, I. Fellegvári, J. Sági, A. Szemzö

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A series of 5-alkyl-, 5-alkenyl- and 5-alkynyl-substituted deoxyuridines and their triphosphate derivatives were synthesized and studied in DNA polymerase reactions. The initial rate of incorporation of the derivatives catalysed by Klenow fragment DNA polymerase enzyme (E. coli) was measured. Calf thymus DNA and synthetic poly (dA-dT) served as templates. The rate values were expressed as a percentage relative to the incorporation rate of natural substrate dTTP. The high-performance liquid chromatographic (HPLC) retention behaviours of the nucleoside derivatives were investigated on silica and reversed-phase stationary phases using various mixtures of ethyl acetate-methanol and methanol-water, as respectively, mobile phases. According to the results of principal component analysis, the HPLC retention data describe the hydrophobic properties of the compounds. The inclusion complex stability constants of the derivatives with cyclodextrins determined by reversed-phase thin-layer chromatography served as a measure of the steric properties of the subtituents. The electronic properties of the 5-substituents were characterized by the Swain-Lupton inductive and resonance parameters. The results of the stepwise linear regression analysis of the nucleotide incorporation rate data and the above-mentioned physico-chemical data revealed the importance of the electronic, steric and hydrophobic properties of the substituents in the DNA polymerase reactions. The importance of the steric parameter was more significant when the poly (dA-dT) template was used instead of the random base sequence template (calf thymus DNA).

Original languageEnglish
Pages (from-to)35-44
Number of pages10
JournalJournal of Chromatography A
Volume506
Issue numberC
DOIs
Publication statusPublished - May 11 1990

Fingerprint

DNA-Directed DNA Polymerase
Poly dA-dT
Nucleotides
Derivatives
Methanol
DNA Polymerase I
Cyclodextrins
Reverse-Phase Chromatography
Thin Layer Chromatography
Principal Component Analysis
Nucleosides
Silicon Dioxide
Thin layer chromatography
Linear Models
Liquids
Regression Analysis
Linear regression
Regression analysis
Electronic properties
Principal component analysis

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry
  • Molecular Medicine

Cite this

Application of chromatographic retention data in an investigation of a quantitative structure-nucleotide incorporation rate relationship. / Valkó, K.; Cserháti, T.; Fellegvári, I.; Sági, J.; Szemzö, A.

In: Journal of Chromatography A, Vol. 506, No. C, 11.05.1990, p. 35-44.

Research output: Contribution to journalArticle

@article{83b960b104304642963bd88a6017e35e,
title = "Application of chromatographic retention data in an investigation of a quantitative structure-nucleotide incorporation rate relationship",
abstract = "A series of 5-alkyl-, 5-alkenyl- and 5-alkynyl-substituted deoxyuridines and their triphosphate derivatives were synthesized and studied in DNA polymerase reactions. The initial rate of incorporation of the derivatives catalysed by Klenow fragment DNA polymerase enzyme (E. coli) was measured. Calf thymus DNA and synthetic poly (dA-dT) served as templates. The rate values were expressed as a percentage relative to the incorporation rate of natural substrate dTTP. The high-performance liquid chromatographic (HPLC) retention behaviours of the nucleoside derivatives were investigated on silica and reversed-phase stationary phases using various mixtures of ethyl acetate-methanol and methanol-water, as respectively, mobile phases. According to the results of principal component analysis, the HPLC retention data describe the hydrophobic properties of the compounds. The inclusion complex stability constants of the derivatives with cyclodextrins determined by reversed-phase thin-layer chromatography served as a measure of the steric properties of the subtituents. The electronic properties of the 5-substituents were characterized by the Swain-Lupton inductive and resonance parameters. The results of the stepwise linear regression analysis of the nucleotide incorporation rate data and the above-mentioned physico-chemical data revealed the importance of the electronic, steric and hydrophobic properties of the substituents in the DNA polymerase reactions. The importance of the steric parameter was more significant when the poly (dA-dT) template was used instead of the random base sequence template (calf thymus DNA).",
author = "K. Valk{\'o} and T. Cserh{\'a}ti and I. Fellegv{\'a}ri and J. S{\'a}gi and A. Szemz{\"o}",
year = "1990",
month = "5",
day = "11",
doi = "10.1016/S0021-9673(01)91565-1",
language = "English",
volume = "506",
pages = "35--44",
journal = "Journal of Chromatography",
issn = "0021-9673",
publisher = "Elsevier",
number = "C",

}

TY - JOUR

T1 - Application of chromatographic retention data in an investigation of a quantitative structure-nucleotide incorporation rate relationship

AU - Valkó, K.

AU - Cserháti, T.

AU - Fellegvári, I.

AU - Sági, J.

AU - Szemzö, A.

PY - 1990/5/11

Y1 - 1990/5/11

N2 - A series of 5-alkyl-, 5-alkenyl- and 5-alkynyl-substituted deoxyuridines and their triphosphate derivatives were synthesized and studied in DNA polymerase reactions. The initial rate of incorporation of the derivatives catalysed by Klenow fragment DNA polymerase enzyme (E. coli) was measured. Calf thymus DNA and synthetic poly (dA-dT) served as templates. The rate values were expressed as a percentage relative to the incorporation rate of natural substrate dTTP. The high-performance liquid chromatographic (HPLC) retention behaviours of the nucleoside derivatives were investigated on silica and reversed-phase stationary phases using various mixtures of ethyl acetate-methanol and methanol-water, as respectively, mobile phases. According to the results of principal component analysis, the HPLC retention data describe the hydrophobic properties of the compounds. The inclusion complex stability constants of the derivatives with cyclodextrins determined by reversed-phase thin-layer chromatography served as a measure of the steric properties of the subtituents. The electronic properties of the 5-substituents were characterized by the Swain-Lupton inductive and resonance parameters. The results of the stepwise linear regression analysis of the nucleotide incorporation rate data and the above-mentioned physico-chemical data revealed the importance of the electronic, steric and hydrophobic properties of the substituents in the DNA polymerase reactions. The importance of the steric parameter was more significant when the poly (dA-dT) template was used instead of the random base sequence template (calf thymus DNA).

AB - A series of 5-alkyl-, 5-alkenyl- and 5-alkynyl-substituted deoxyuridines and their triphosphate derivatives were synthesized and studied in DNA polymerase reactions. The initial rate of incorporation of the derivatives catalysed by Klenow fragment DNA polymerase enzyme (E. coli) was measured. Calf thymus DNA and synthetic poly (dA-dT) served as templates. The rate values were expressed as a percentage relative to the incorporation rate of natural substrate dTTP. The high-performance liquid chromatographic (HPLC) retention behaviours of the nucleoside derivatives were investigated on silica and reversed-phase stationary phases using various mixtures of ethyl acetate-methanol and methanol-water, as respectively, mobile phases. According to the results of principal component analysis, the HPLC retention data describe the hydrophobic properties of the compounds. The inclusion complex stability constants of the derivatives with cyclodextrins determined by reversed-phase thin-layer chromatography served as a measure of the steric properties of the subtituents. The electronic properties of the 5-substituents were characterized by the Swain-Lupton inductive and resonance parameters. The results of the stepwise linear regression analysis of the nucleotide incorporation rate data and the above-mentioned physico-chemical data revealed the importance of the electronic, steric and hydrophobic properties of the substituents in the DNA polymerase reactions. The importance of the steric parameter was more significant when the poly (dA-dT) template was used instead of the random base sequence template (calf thymus DNA).

UR - http://www.scopus.com/inward/record.url?scp=0025339961&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025339961&partnerID=8YFLogxK

U2 - 10.1016/S0021-9673(01)91565-1

DO - 10.1016/S0021-9673(01)91565-1

M3 - Article

AN - SCOPUS:0025339961

VL - 506

SP - 35

EP - 44

JO - Journal of Chromatography

JF - Journal of Chromatography

SN - 0021-9673

IS - C

ER -