title: ÐÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ðµ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½ÑÑ ÑÐ²Ð¾Ð¹ÑÑÐ² nadh-adh Ð² ÑÐ¼ÐµÑÐ¸ ÐµÑÑÐµÑÑÐ²ÐµÐ½Ð½ÑÑ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½ÑÑ ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑÐ¾Ð² Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾Ð¹ Ð¿Ð¾Ð»ÑÑÐ¸Ð·Ð°ÑÐ¸Ð¾Ð½Ð½Ð¾Ð¹ ÑÐ¿ÐµÐºÑÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ð¸ Ñ Ð²ÑÑÐ¾ÐºÐ¸Ð¼ Ð²ÑÐµÐ¼ÐµÐ½Ð½ÑÐ¼ ÑÐ°Ð·ÑÐµÑÐµÐ½Ð¸ÐµÐ¼
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Investigation of fluorescent properties of NADH-ADH complex in presence of natural enzymes by means of time-resolved polarization spectroscopy
Creators: name: Ð¯ÑÐºÐ¾Ð², ÐÐ¼Ð¸ÑÑÐ¸Ð¹
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2022
Subjects: ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ñ
NADH
ADH
Ð¿Ð¾Ð»ÑÑÐ¸Ð·Ð°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ¿ÐµÐºÑÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ñ
Ð²ÑÐµÐ¼Ñ-ÑÐ°Ð·ÑÐµÑÐµÐ½Ð½Ð°Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ñ
fluorescence
polarization spectroscopy
time-resolved fluorescence

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑÐ° NADH-ADH, Ð¿ÑÐ¸ Ð´Ð²ÑÑÑÐ¾ÑÐ¾Ð½Ð½Ð¾Ð¼ Ð²Ð¾Ð·Ð±ÑÐ¶Ð´ÐµÐ½Ð¸Ð¸ ÑÐµÐ¼ÑÐ¾ÑÐµÐºÑÐ½Ð´Ð½ÑÐ¼Ð¸ Ð»Ð°Ð·ÐµÑÐ½ÑÐ¼Ð¸ Ð¸Ð¼Ð¿ÑÐ»ÑÑÐ°Ð¼Ð¸ Ñ Ð´Ð»Ð¸Ð½Ð½Ð¾Ð¹ Ð²Ð¾Ð»Ð½Ñ 720 Ð¸ 840 Ð½Ð¼. Ð¦ÐµÐ»ÑÑ Ð½Ð°ÑÑÐ¾ÑÑÐµÐ¹ ÑÐ°Ð±Ð¾ÑÑ ÑÐ²Ð»ÑÐµÑÑÑ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ ÑÐ¸Ð³Ð½Ð°Ð»Ð° ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ ÐºÐ¾ÑÐµÑÐ¼ÐµÐ½ÑÐ° NADH, Ð½Ð°ÑÐ¾Ð´ÑÑÐµÐ³Ð¾ÑÑ Ð² ÑÐ¼ÐµÑÐ¸ ÐµÑÑÐµÑÑÐ²ÐµÐ½Ð½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð². ÐÐ»Ñ Ð¿Ð¾Ð»ÑÑÐµÐ½Ð¸Ñ ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÐ°Ð»ÑÐ½ÑÑ ÑÐ¸Ð³Ð½Ð°Ð»Ð¾Ð² Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð»Ð°ÑÑ ÑÐ¸ÑÑÐµÐ¼Ð° ÑÑÐµÑÐ° ÐµÐ´Ð¸Ð½Ð¸ÑÐ½ÑÑ ÑÐ¾ÑÐ¾Ð½Ð¾Ð² Ñ Ð²ÑÐµÐ¼ÐµÐ½Ð½Ð¾Ð¹ ÐºÐ¾ÑÑÐµÐ»ÑÑÐ¸ÐµÐ¹ (TCSPC). Ð ÑÐ°Ð¼ÐºÐ°Ñ Ð´Ð°Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¸ Ð¿ÑÐ¾Ð°Ð½Ð°Ð»Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°Ð½Ñ ÑÐ¿ÐµÐºÑÑÑ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ ÑÐ°ÑÑÐ²Ð¾ÑÐ° ADH, Ð¿Ð¾Ð»ÑÑÐµÐ½Ð½ÑÑ Ð¿ÑÐ¸ Ð´Ð²ÑÑÑÐ¾ÑÐ¾Ð½Ð½Ð¾Ð¼ Ð²Ð¾Ð·Ð±ÑÐ¶Ð´ÐµÐ½Ð¸Ð¸ Ð½Ð° Ð´Ð»Ð¸Ð½Ð°Ñ Ð²Ð¾Ð»Ð½ 720 Ð¸ 840 Ð½Ð¼. ÐÐ»Ñ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° ÑÐ¸Ð¼Ð¸ÑÐµÑÐºÐ¾Ð³Ð¾ ÑÐ¾ÑÑÐ°Ð²Ð° ÑÐ¼ÐµÑÐ¸ Ð±ÐµÐ»ÐºÐ¾Ð² Ð±ÑÐ»Ð° Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° Ð¾ÑÐ¸ÑÑÐºÐ° ÑÐ°ÑÑÐ²Ð¾ÑÐ° Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ð¸, Ð² ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ ÐºÐ¾ÑÐ¾ÑÐ¾Ð¹ Ð¾Ð½ Ð±ÑÐ» ÑÐ°Ð·Ð´ÐµÐ»ÐµÐ½ Ð½Ð° ÑÑÐ°ÐºÑÐ¸Ð¸, Ð¸ Ð¿Ð¾ÑÐ»ÐµÐ´ÑÑÑÐ¸Ð¹ Ð°Ð½Ð°Ð»Ð¸Ð· ÑÑÐ°ÐºÑÐ¸Ð¹ Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ Ð¼Ð°ÑÑ-ÑÐ¿ÐµÐºÑÑÐ¾Ð¼ÐµÑÑÐ¸Ð¸. ÐÐ»Ñ ÑÑÐ°ÐºÑÐ¸Ð¹, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ñ Ð¸ÑÑÐ»ÐµÐ´ÑÐµÐ¼ÑÐ¹ ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑ NADH-ADH, Ð±ÑÐ»Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ñ Ð¸Ð·Ð¼ÐµÑÐµÐ½Ð¸Ñ ÑÐ¸Ð³Ð½Ð°Ð»Ð¾Ð² Ð·Ð°ÑÑÑÐ°Ð½Ð¸Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ ÑÐ¿ÐµÐºÑÑÐ°Ð»ÑÐ½ÑÑ Ð´Ð¸Ð°Ð¿Ð°Ð·Ð¾Ð½Ð°Ñ 426-446 Ð½Ð¼ Ð¸ 460-480 Ð½Ð¼ Ð¿ÑÐ¸ Ð´Ð²ÑÑÑÐ¾ÑÐ¾Ð½Ð½Ð¾Ð¼ Ð²Ð¾Ð·Ð±ÑÐ¶Ð´ÐµÐ½Ð¸Ð¸ Ð½Ð° Ð´Ð»Ð¸Ð½Ð°Ñ Ð²Ð¾Ð»Ð½ 720 Ð¸ 840 Ð½Ð¼. ÐÐ¾Ð»ÑÑÐµÐ½Ð½ÑÐµ ÑÐ¸Ð³Ð½Ð°Ð»Ñ Ð·Ð°ÑÑÑÐ°Ð½Ð¸Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ Ð±ÑÐ»Ð¸ Ð¿ÑÐ¾Ð°Ð½Ð°Ð»Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°Ð½Ñ Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ ÑÑÐ½ÐºÑÐ¸Ð¹ ÑÑÐ¼Ð¼Ñ 2-Ñ, 3-Ñ Ð¸ 4-Ñ ÑÐºÑÐ¿Ð¾Ð½ÐµÐ½Ñ, Ð² ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ ÑÐµÐ³Ð¾ Ð±ÑÐ»Ð¸ Ð¾Ð¿ÑÐµÐ´ÐµÐ»ÐµÐ½Ñ ÑÐ»ÐµÐ´ÑÑÑÐ¸Ðµ Ð¿Ð°ÑÐ°Ð¼ÐµÑÑÑ: Ð²ÑÐµÐ¼ÐµÐ½Ð° Ð¶Ð¸Ð·Ð½Ð¸ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ Ð¸ ÑÐ¾Ð¾ÑÐ²ÐµÑÑÑÐ²ÑÑÑÐ¸Ðµ Ð¸Ð¼Â Ð²ÐµÑÐ¾Ð²ÑÐµ ÐºÐ¾ÑÑÑÐ¸ÑÐ¸ÐµÐ½ÑÑ. ÐÑÐ»Ð¾ Ð¿ÑÐ¾Ð¸Ð·Ð²ÐµÐ´ÐµÐ½Ð¾ ÑÑÐ°Ð²Ð½ÐµÐ½Ð¸Ðµ Ð¼Ð¾Ð´ÐµÐ»ÐµÐ¹ Ð°Ð¿Ð¿ÑÐ¾ÐºÑÐ¸Ð¼Ð°ÑÐ¸Ð¸, Ð² ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ ÑÐµÐ³Ð¾ Ð±ÑÐ»Ð° Ð²ÑÐ±ÑÐ°Ð½Ð° ÑÑÐ½ÐºÑÐ¸Ñ, Ð½Ð°Ð¸Ð»ÑÑÑÐ¸Ð¼ Ð¾Ð±ÑÐ°Ð·Ð¾Ð¼ Ð¾Ð¿Ð¸ÑÑÐ²Ð°ÑÑÐ°Ñ ÑÐ¸Ð³Ð½Ð°Ð»Ñ Ð·Ð°ÑÑÑÐ°Ð½Ð¸Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ Ð¸ÑÑÐ»ÐµÐ´ÑÐµÐ¼ÑÑ ÑÑÐ°ÐºÑÐ¸Ð¹. Ð ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð½ÑÑ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ð¹ Ð¸ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° Ð¿Ð¾Ð»ÑÑÐµÐ½Ð½ÑÑ Ð´Ð°Ð½Ð½ÑÑ Ð±ÑÐ»Ð¸ Ð¾Ð±Ð½Ð°ÑÑÐ¶ÐµÐ½Ñ Ð¸ Ð¸Ð´ÐµÐ½ÑÐ¸ÑÐ¸ÑÐ¸ÑÐ¾Ð²Ð°Ð½Ñ Ð¿ÑÐ¸Ð¼ÐµÑÐ½ÑÐµ Ð±ÐµÐ»ÐºÐ¸, Ð½Ð°ÑÐ¾Ð´ÑÑÐ¸ÐµÑÑ Ð² ÑÐ°ÑÑÐ²Ð¾ÑÐµ ADH, Ð° ÑÐ°ÐºÐ¶Ðµ ÑÑÐ¿ÐµÑÐ½Ð¾ Ð²ÑÐ´ÐµÐ»ÐµÐ½ ÑÐ¸Ð³Ð½Ð°Ð» ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑÐ° NADH-ADH Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ ÑÐ·ÐºÐ¾Ð¿Ð¾Ð»Ð¾ÑÐ½Ð¾Ð³Ð¾ Ð¸Ð½ÑÐµÑÑÐµÑÐµÐ½ÑÐ¸Ð¾Ð½Ð½Ð¾Ð³Ð¾ ÑÐ¸Ð»ÑÑÑÐ°.
description: This work is devoted to the study of the fluorescence of the NADH-ADH complex, under two-photon excitation by femtosecond laser pulses with a wavelength of 720 and 840 nm. The aim of this work is to isolate the fluorescence signal of the biological coenzyme NADH, which is in a mixture of natural proteins. A time-correlated single photon counting system (TCSPC) was used to obtain experimental signals. Within the framework of this work, the fluorescence spectra of the ADH solution at excitation wavelengths of 720 and 840 nm were analyzed. Then the solution was cleaned, as a result of which it was divided into fractions, and subsequent analysis by chromatography and mass spectrometry, respectively. Further, fluorescence attenuation signals of certain fractions were measured at excitation wavelengths of 720 and 840 nm, in the selected fluorescence spectral ranges of 426-446 nm and 460-480 nm. The obtained fluorescence attenuation signals were analyzed using the sum functions of 2, 3 and 4 exponents, as a result of which the following parameters were calculated: fluorescence lifetime and corresponding weighting coefficients. The approximation models were compared to select the best number of exponents for each of the fraction. As a result of the studies and analysis of the data obtained, impurity proteins in the ADH solution were detected and identified, and the NADH fluorescence signal was successfully isolated using a bandpass filter.