Efficient production of highly purified Late Embryogenesis Abundant (LEA) protein from Arabidopsis thaliana by recombinant DNA technology
Аутори
Marković, NemanjaMilić Komić, Sonja
Radosavljević, Jelena
Pantelić, Ana
Kilibarda, Nataša
Vidović, Marija
Остала ауторства
Spasojević, IvanКонференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Late embryogenesis abundant (LEA) proteins are induced in cellular dehydration, such as
freezing, drought, or desiccation. They can be involved in antioxidative defense, ion
sequestration, and structural stabilization of both membranes and enzymes during freezing
or drying, while by forming intracellular proteinaceous condensates they increase
structural integrity and intracellular viscosity of cells during desiccation 1. The genome of
the model plant Arabidopsis thaliana contains 51 genes encoding LEA proteins2. The
majority of these LEA proteins (35%) belongs to Pfam LEA_4 (PF02987) family. In silico
analysis suggested that these proteins are highly hydrophilic proteins with significant
intrinsically disordered protein (IDP) properties. In order to evaluate structural properties
and possible functions of LEA_4 protein family under different water content, a
representative AtLEA25 protein (At2g42560, 635 aa), naturally located in the cytoplasm
of seeds3 was obtained in Esch...erichia coli by recombinant DNA technology. Although this
technology has been traditionally used to over-express and purify various globular
proteins, numerous reports have shown that the IDPs, due to their structural plasicity are
naturally highly susceptible to proteolytic cleavage. To conduct structural and functional
studies we developed a robust method to produce highly purified (>95% pure) AtLEA25
with no detectable amount of protein breakdown products.
Кључне речи:
LEA4 protein family / AtLEA25 / Escherichia coli / intrinsically disordered proteins / Protein purificationИзвор:
Biochemical Insights into Molecular Mechanisms, 2021, 98-99Издавач:
- Belgrade : Serbian Biochemical Society
Финансирање / пројекти:
- LEAPSyn-SCI - Late Embryogenesis Abundant Proteins: Structural Characterisation and Interaction With Α-Synuclein (RS-ScienceFundRS-Promis-6039663)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200053 (Универзитет у Београду, Институт за мултидисциплинарна истраживања) (RS-MESTD-inst-2020-200053)
Напомена:
- Abstract: Serbian Biochemical Society, 10th Conference, Kragujevac, Serbia, 24.09.2021
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Marković, Nemanja AU - Milić Komić, Sonja AU - Radosavljević, Jelena AU - Pantelić, Ana AU - Kilibarda, Nataša AU - Vidović, Marija PY - 2021 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/1870 AB - Late embryogenesis abundant (LEA) proteins are induced in cellular dehydration, such as freezing, drought, or desiccation. They can be involved in antioxidative defense, ion sequestration, and structural stabilization of both membranes and enzymes during freezing or drying, while by forming intracellular proteinaceous condensates they increase structural integrity and intracellular viscosity of cells during desiccation 1. The genome of the model plant Arabidopsis thaliana contains 51 genes encoding LEA proteins2. The majority of these LEA proteins (35%) belongs to Pfam LEA_4 (PF02987) family. In silico analysis suggested that these proteins are highly hydrophilic proteins with significant intrinsically disordered protein (IDP) properties. In order to evaluate structural properties and possible functions of LEA_4 protein family under different water content, a representative AtLEA25 protein (At2g42560, 635 aa), naturally located in the cytoplasm of seeds3 was obtained in Escherichia coli by recombinant DNA technology. Although this technology has been traditionally used to over-express and purify various globular proteins, numerous reports have shown that the IDPs, due to their structural plasicity are naturally highly susceptible to proteolytic cleavage. To conduct structural and functional studies we developed a robust method to produce highly purified (>95% pure) AtLEA25 with no detectable amount of protein breakdown products. PB - Belgrade : Serbian Biochemical Society C3 - Biochemical Insights into Molecular Mechanisms T1 - Efficient production of highly purified Late Embryogenesis Abundant (LEA) protein from Arabidopsis thaliana by recombinant DNA technology EP - 99 SP - 98 UR - https://hdl.handle.net/21.15107/rcub_imagine_1870 ER -
@conference{ author = "Marković, Nemanja and Milić Komić, Sonja and Radosavljević, Jelena and Pantelić, Ana and Kilibarda, Nataša and Vidović, Marija", year = "2021", abstract = "Late embryogenesis abundant (LEA) proteins are induced in cellular dehydration, such as freezing, drought, or desiccation. They can be involved in antioxidative defense, ion sequestration, and structural stabilization of both membranes and enzymes during freezing or drying, while by forming intracellular proteinaceous condensates they increase structural integrity and intracellular viscosity of cells during desiccation 1. The genome of the model plant Arabidopsis thaliana contains 51 genes encoding LEA proteins2. The majority of these LEA proteins (35%) belongs to Pfam LEA_4 (PF02987) family. In silico analysis suggested that these proteins are highly hydrophilic proteins with significant intrinsically disordered protein (IDP) properties. In order to evaluate structural properties and possible functions of LEA_4 protein family under different water content, a representative AtLEA25 protein (At2g42560, 635 aa), naturally located in the cytoplasm of seeds3 was obtained in Escherichia coli by recombinant DNA technology. Although this technology has been traditionally used to over-express and purify various globular proteins, numerous reports have shown that the IDPs, due to their structural plasicity are naturally highly susceptible to proteolytic cleavage. To conduct structural and functional studies we developed a robust method to produce highly purified (>95% pure) AtLEA25 with no detectable amount of protein breakdown products.", publisher = "Belgrade : Serbian Biochemical Society", journal = "Biochemical Insights into Molecular Mechanisms", title = "Efficient production of highly purified Late Embryogenesis Abundant (LEA) protein from Arabidopsis thaliana by recombinant DNA technology", pages = "99-98", url = "https://hdl.handle.net/21.15107/rcub_imagine_1870" }
Marković, N., Milić Komić, S., Radosavljević, J., Pantelić, A., Kilibarda, N.,& Vidović, M.. (2021). Efficient production of highly purified Late Embryogenesis Abundant (LEA) protein from Arabidopsis thaliana by recombinant DNA technology. in Biochemical Insights into Molecular Mechanisms Belgrade : Serbian Biochemical Society., 98-99. https://hdl.handle.net/21.15107/rcub_imagine_1870
Marković N, Milić Komić S, Radosavljević J, Pantelić A, Kilibarda N, Vidović M. Efficient production of highly purified Late Embryogenesis Abundant (LEA) protein from Arabidopsis thaliana by recombinant DNA technology. in Biochemical Insights into Molecular Mechanisms. 2021;:98-99. https://hdl.handle.net/21.15107/rcub_imagine_1870 .
Marković, Nemanja, Milić Komić, Sonja, Radosavljević, Jelena, Pantelić, Ana, Kilibarda, Nataša, Vidović, Marija, "Efficient production of highly purified Late Embryogenesis Abundant (LEA) protein from Arabidopsis thaliana by recombinant DNA technology" in Biochemical Insights into Molecular Mechanisms (2021):98-99, https://hdl.handle.net/21.15107/rcub_imagine_1870 .