Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
Expression of Recombinant LDLR–EGFP Fusion Protein in HEK-293 Cells as a Promising Tool to Assess the Effect of LDLR Gene Mutations. / Polyakov, D. S.; Grudinina, N. A.; Bogoslovskaya, T. Yu; Sokolov, A. V.; Mandelshtam, M. Yu; Vasilyev, V. B.
в: Cell and Tissue Biology, Том 12, № 2, 2018, стр. 153-159.Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
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TY - JOUR
T1 - Expression of Recombinant LDLR–EGFP Fusion Protein in HEK-293 Cells as a Promising Tool to Assess the Effect of LDLR Gene Mutations
AU - Polyakov, D. S.
AU - Grudinina, N. A.
AU - Bogoslovskaya, T. Yu
AU - Sokolov, A. V.
AU - Mandelshtam, M. Yu
AU - Vasilyev, V. B.
N1 - Polyakov, D.S., Grudinina, N.A., Bogoslovskaya, T.Y. et al. Expression of Recombinant LDLR–EGFP Fusion Protein in HEK-293 Cells as a Promising Tool to Assess the Effect of LDLR Gene Mutations. Cell Tiss. Biol. 12, 153–159 (2018). https://doi.org/10.1134/S1990519X18020098
PY - 2018
Y1 - 2018
N2 - Mutations in the low density lipoprotein receptor gene (LDLR) frequently impair folding and intracellular traffic of the receptor protein, resulting in the development of a monogenic disorder, familial hypercholesterolemia (FH). Identification of novel LDLR mutations requires confirmation of their functional importance in distinguishing pathogenic mutations from neutral changes in the aminoacid sequence. To elaborate a system for evaluation of the effect of mutation on the folding and intracellular transport of the LDLR, as well as its ability to bind low density lipoprotein (LDL), we constructed a plasmid containing LDLR cDNA and the gene of enhanced green fluorescent protein (EGFP). Confocal microscopy has shown that, upon transient transfection of HEK293 cells with the plasmid, the recombinant fusion protein LDLR–EGFP is transported onto the cellular membrane and binds labeled LDL. This construct will be further modified by site-directed mutagenesis to reproduce the LDLR missense mutations most common in the population of northwest Russia so as to study the subcellular localization and function of the modified chimeric protein.
AB - Mutations in the low density lipoprotein receptor gene (LDLR) frequently impair folding and intracellular traffic of the receptor protein, resulting in the development of a monogenic disorder, familial hypercholesterolemia (FH). Identification of novel LDLR mutations requires confirmation of their functional importance in distinguishing pathogenic mutations from neutral changes in the aminoacid sequence. To elaborate a system for evaluation of the effect of mutation on the folding and intracellular transport of the LDLR, as well as its ability to bind low density lipoprotein (LDL), we constructed a plasmid containing LDLR cDNA and the gene of enhanced green fluorescent protein (EGFP). Confocal microscopy has shown that, upon transient transfection of HEK293 cells with the plasmid, the recombinant fusion protein LDLR–EGFP is transported onto the cellular membrane and binds labeled LDL. This construct will be further modified by site-directed mutagenesis to reproduce the LDLR missense mutations most common in the population of northwest Russia so as to study the subcellular localization and function of the modified chimeric protein.
KW - familial hypercholesterolemia
KW - fusion protein
KW - gene expression
KW - green fluorescent protein
KW - low density lipoprotein receptor
UR - http://www.scopus.com/inward/record.url?scp=85045645800&partnerID=8YFLogxK
U2 - 10.1134/S1990519X18020098
DO - 10.1134/S1990519X18020098
M3 - Article
AN - SCOPUS:85045645800
VL - 12
SP - 153
EP - 159
JO - Cell and Tissue Biology
JF - Cell and Tissue Biology
SN - 1990-519X
IS - 2
ER -
ID: 42247378