Acta NaturaeActa NaturaeActa Naturae2075-8251Acta Naturae Ltd2764510.32607/actanaturae.27645Research ArticlesЭкспериментальные статьиResearch ArticleTest system for studying biotin transport upon SLC5A6 gene inactivationТест-система для изучения транспорта биотина при инактивации гена SLC5A6RudenkoA. Yu.РуденкоА. Ю.
Russian Federation

Belozersky Institute of Physico-Chemical Biology; Faculty of Fundamental Medicine

Институт физико-химической биологии имени А.Н. Белозерского; факультет фундаментальной медицины

RudenkoAY@my.msu.ruZotovaP. A.ЗотоваП. А.
Russian Federation

Belozersky Institute of Physico-Chemical Biology; Department of Chemistry

Институт физико-химической биологии имени А.Н. Белозерского; химический факультет

RudenkoAY@my.msu.ruAverinaO. A.АверинаО. А.
Russian Federation

Belozersky Institute of Physico-Chemical Biology

Институт физико-химической биологии имени А.Н. Белозерского

RudenkoAY@my.msu.ruPriymakA. V.ПриймакА. В.
Russian Federation

Belozersky Institute of Physico-Chemical Biology; Институт физико-химической биологии имени А.Н. Белозерского

Faculty of Fundamental Medicine; факультет фундаментальной медицины

RudenkoAY@my.msu.ruRubtsovaM. P.РубцоваМ. П.
Russian Federation

Department of Chemistry, Lomonosov Moscow State University

химический факультет, Московский государственный университет имени М.В. Ломоносова

RudenkoAY@my.msu.ruMariasinaS. S.МарьясинаС. С.
Russian Federation

Belozersky Institute of Physico-Chemical Biology; Faculty of Fundamental Medicine Lomonosov Moscow State University; Department of Chemistry, Lomonosov Moscow State University

Институт физико-химической биологии имени А.Н. Белозерского; факультет фундаментальной медицины Московский государственный университет имени М.В. Ломоносова; химический факультет Московский государственный университет имени М.В. Ломоносова

RudenkoAY@my.msu.ruOzhiganovR. M.ОжигановР. М.
Russian Federation

Belozersky Institute of Physico-Chemical Biology

Институт физико-химической биологии имени А.Н. Белозерского

RudenkoAY@my.msu.ruDontsovaO. A.ДонцоваО. А.
Russian Federation

Belozersky Institute of Physico-Chemical Biology; Department of Chemistry, Lomonosov Moscow State Universit

Институт физико-химической биологии имени А.Н. Белозерского; химический факультет, Московский государственный университет имени М.В. Ломоносова

RudenkoAY@my.msu.ruSergievP. V.СергиевП. В.
Russian Federation

Belozersky Institute of Physico-Chemical Biology; Department of Chemistry, Lomonosov Moscow State University

Институт физико-химической биологии имени А.Н. Белозерского; химический факультет, Московский государственный университет имени М.В. Ломоносова

RudenkoAY@my.msu.ruLomonosov Moscow State UniversityМосковский государственный университет имени М.В. ЛомоносоваShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of SciencesГосударственный научный центр Институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова РАНResearch and Educational Resource Center “Pharmacy”, RUDN UniversityИнститут фармации и биотехнологии РУДНHigher Chemical College RAS, Mendeleev University of Chemical TechnologyРоссийский химико-технологический университет имени Д.И. Менделеева, Высший химический колледж Российской академии наукCenter for Molecular and Cellular Biology, Skolkovo Institute of Science and TechnologyЦентр молекулярной и клеточной биологии, Сколтех141020251731191292702202521052025Copyright ©; 2025, Rudenko A.Y., Zotova P.A., Averina O.A., Priymak A.V., Rubtsova M.P., Mariasina S.S., Ozhiganov R.M., Dontsova O.A., Sergiev P.V.Copyright ©; 2025, Руденко А.Ю., Зотова П.А., Аверина О.А., Приймак А.В., Рубцова М.П., Марьясина С.С., Ожиганов Р.М., Донцова О.А., Сергиев П.В.2025Rudenko A.Y., Zotova P.A., Averina O.A., Priymak A.V., Rubtsova M.P., Mariasina S.S., Ozhiganov R.M., Dontsova O.A., Sergiev P.V.Руденко А.Ю., Зотова П.А., Аверина О.А., Приймак А.В., Рубцова М.П., Марьясина С.С., Ожиганов Р.М., Донцова О.А., Сергиев П.В.https://creativecommons.org/licenses/by/4.0https://actanaturae.ru/2075-8251/article/view/27645

This paper introduces a test system for the investigation of biotin transport following inactivation of the SLC5A6 gene, which encodes the sodium-dependent multivitamin transporter SLC5A6. The aim was to develop a method for assessing the efficiency of biotin penetration across the cell membrane following inactivation of the SLC5A6 gene and to explore the feasibility of delivering biotin derivatives into cells independent of SLC5A6. The test system is built upon modified HEK293 cell lines with overexpression of the BirA* biotin ligase, with the first line comprising a functional SLC5A6 gene and the second one involving an inactivated version of this gene mimicking impaired biotin transport. This test system was used to investigate the transport of biotin and its two derivatives, namely the biotin conjugate with p-aminophenylalanine (Bio-1) and biotin methyl ester (Bio-2), through the cell membrane. It has been determined that biotin and its methyl ester (Bio-2) can enter cells independently of the SLC5A6 transporter, which points to the presence of alternative transport pathways. The biotin derivative Bio-1, which contains p-aminophenylalanine, is internalized into cells solely through the hSMVT transporter. The novel test system will serve as a tool for investigating the pathways involved in vitamin entry into cells and for developing therapeutic strategies for individuals with mutations in the SLC5A6 gene, as well as other transport-related genes.

Представлена тест-система для изучения транспорта биотина при инактивации гена SLC5A6, кодирующего натрий-зависимый поливитаминный транспортер SLC5A6. Цель работы состоит в разработке метода оценки эффективности проникновения биотина через клеточную мембрану при инактивации гена SLС5A6, а также в изучении возможности SLC5A6-независимой доставки производных биотина в клетки. Тест-система основана на использовании модифицированных клеточных линий HEK293, сверхэкспрессирующих биотинлигазу BirA*. Одна линия содержит функциональный ген SLC5A6, в другой линии этот ген инактивирован для имитации нарушения транспорта биотина. С использованием разработанной тест-системы изучен транспорт через клеточную мембрану биотина, а также двух его производных: конъюгата биотина с п-аминофенилаланином (Bio-1) и метилового эфира биотина (Bio-2). Установлено, что биотин и его метиловый эфир (Bio-2) способны проникать в клетки независимо от транспортера SLC5A6, что указывает на существование альтернативных путей транспорта. Производное биотина с п-аминофенилаланином (Bio-1) проникает в клетки только через hSMVT. Разработанная тест-система является новым инструментом для изучения путей поступления витаминов в клетки и разработки терапевтических стратегий для пациентов с мутацией гена SLC5A6, а также генов других транспортеров.

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