Oligomeric state investigation of flavocytochrome CYP102A1 using afm with standard and supersharp probes

Abstract

Atomic force microscopy with two types of probes – standard (radius of curvature R~10 nm) and supersharp (R~2 nm) – was used to determine CYP102A1oligomeric state. CYP102A1 images were obtained in a liquid, air and vacuum environment using the standard probes, also a ratio of monomers to oligomers (a) of CYP102A1 were determined as a=0.48:0.52. At the same time use of standard probes did not allow to resolve the structure of these oligomers. Supersharp probes allowed to obtain the data about the monomers to oligomers ratio, and also about the dimers/trimers/tetramers ratio in air and vacuum. So, a ratio a of CYP102A1 in liquid can be determined by the standard probes, and an oligomeric state of protein can be specified by the supersharp probes.

Atomic Force Microscopy for Protein Detection and Their Physicoсhemical Characterization

“…The information on the enzyme’s structure is very important for elucidation of the reaction mechanisms of (I) and (II) groups. AFM was capable of visualizing and obtaining the height of protein molecules and protein-protein complexes forming in those systems [ 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 ] in a tapping mode. Analysis of the density distribution with height ρ ( h ) and its approximation allowed for the determination of parameters of single proteins as well as protein complexes.…”

Monitoring of microwave emission of HRP system during the enzyme functioning

“…That is why this system is often used as an object in enzymatic reaction studies [10] , [11] , [12] , [13] , [14] and was chosen in the present work to investigate the microwave emission. Besides, in our earlier study on the same it was shown by atomic force microscopy that the functioning of the enzyme is accompanied by fluctuations of the protein globule [12] , [13] , [14] , [15] , [16] . These fluctuations are able to cause disturbances of aqueous environment of the cytochrome CYP102 A1 molecules thereby generating the microwave emission from the whole solution.…”

AFM fishing of proteins under impulse electric field

“…Комбинация АСМ-фишинга с МС анализом имеет большие потенциальные возможности для медицинской протеомики. Это связано с тем, что АСМ-фишинг позволяет одновременно вылавливать, концентрировать и визуализировать низкокопийные белки и их комплексы [4][5][6][7][8], а высокочувствительный МС анализ позволяет идентифицировать выловленные белки. С помощью АСМ-фишинга возможна регистрация белков в диапазоне концентраций 10 -14 -10 -16 M [2].…”