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Characterization of TELSAM Chaperone...

Gajjar, Parag Laljibhai.

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Characterization of TELSAM Chaperone Linker to Improve Crystallization Resolution and Structural Elucidation of the Hydrogenase Maturase Complex.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Characterization of TELSAM Chaperone Linker to Improve Crystallization Resolution and Structural Elucidation of the Hydrogenase Maturase Complex./
作者:	Gajjar, Parag Laljibhai.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2023,
面頁冊數:	110 p.
附註:	Source: Dissertations Abstracts International, Volume: 84-11, Section: B.
Contained By:	Dissertations Abstracts International84-11B.
標題:	Leukemia. -
電子資源:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30396752
ISBN:	9798379470975

Characterization of TELSAM Chaperone Linker to Improve Crystallization Resolution and Structural Elucidation of the Hydrogenase Maturase Complex.
Gajjar, Parag Laljibhai.

Characterization of TELSAM Chaperone Linker to Improve Crystallization Resolution and Structural Elucidation of the Hydrogenase Maturase Complex. - Ann Arbor : ProQuest Dissertations & Theses, 2023 - 110 p.

Source: Dissertations Abstracts International, Volume: 84-11, Section: B.

Thesis (Ph.D.)--Brigham Young University, 2023.

TELSAM crystallization can be a revolutionary tool for the facile crystallization of proteins. TELSAM can increase the rate of crystallization and form crystals at low protein concentrations without direct contact between TELSAM polymers and, in some cases, with very minimal crystal contacts overall. To further understand and characterize TELSAM crystallization, we sought to understand the requirements for the composition of the linker confirmation between TELSAM and the fused target protein. We evaluated four linkers, AlaAla, AlaVal, ThrVal, and ThrThr, between 1TEL and the human CMG2 vWa domain. We compared the number of successful crystallization conditions, the number of crystals, the average and best diffraction resolution, and the refinement parameters for the above constructs. We also tested the effect of the fusion protein SUMO on crystallization. We discovered that rigidification of the linker can improve diffraction resolution, likely by decreasing the possible orientations of the vWa domains in the crystal and that omitting the SUMO domain from the construct also improves the diffraction resolution.The hydrogenase enzyme is involved in the formation and degradation of hydrogen in various biochemical reactions in multiple organisms. It represents a promising avenue for biohydrogen production in the future. The hydrogenase uses a unique iron-sulfur cluster (termed the H-cluster) to generate hydrogen. Three maturase enzymes: HydF (~46 KDa), HydE (~40KDa), and HydG (~55KDa), are involved in the H-cluster assembly/maturation process. Structural characterization of complexes of the maturase enzymes is lacking and needed to better understand the H-cluster assembly. We have expressed and purified the hydrogenase maturase complex. Sizeexclusion chromatography supports the existence of a ~175 KDa complex containing dimeric HydF and monomeric HydG and HydE. We have initial negative stain 2D class averages and a low-resolution 3D reconstruction of the maturation complex. Our next step will be to determine a high-resolution cryo-EM structure of the maturase complex.

ISBN: 9798379470975Subjects--Topical Terms:

677274
Leukemia.

Characterization of TELSAM Chaperone Linker to Improve Crystallization Resolution and Structural Elucidation of the Hydrogenase Maturase Complex.
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520 $a TELSAM crystallization can be a revolutionary tool for the facile crystallization of proteins. TELSAM can increase the rate of crystallization and form crystals at low protein concentrations without direct contact between TELSAM polymers and, in some cases, with very minimal crystal contacts overall. To further understand and characterize TELSAM crystallization, we sought to understand the requirements for the composition of the linker confirmation between TELSAM and the fused target protein. We evaluated four linkers, AlaAla, AlaVal, ThrVal, and ThrThr, between 1TEL and the human CMG2 vWa domain. We compared the number of successful crystallization conditions, the number of crystals, the average and best diffraction resolution, and the refinement parameters for the above constructs. We also tested the effect of the fusion protein SUMO on crystallization. We discovered that rigidification of the linker can improve diffraction resolution, likely by decreasing the possible orientations of the vWa domains in the crystal and that omitting the SUMO domain from the construct also improves the diffraction resolution.The hydrogenase enzyme is involved in the formation and degradation of hydrogen in various biochemical reactions in multiple organisms. It represents a promising avenue for biohydrogen production in the future. The hydrogenase uses a unique iron-sulfur cluster (termed the H-cluster) to generate hydrogen. Three maturase enzymes: HydF (~46 KDa), HydE (~40KDa), and HydG (~55KDa), are involved in the H-cluster assembly/maturation process. Structural characterization of complexes of the maturase enzymes is lacking and needed to better understand the H-cluster assembly. We have expressed and purified the hydrogenase maturase complex. Sizeexclusion chromatography supports the existence of a ~175 KDa complex containing dimeric HydF and monomeric HydG and HydE. We have initial negative stain 2D class averages and a low-resolution 3D reconstruction of the maturation complex. Our next step will be to determine a high-resolution cryo-EM structure of the maturase complex.
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650 4 $a Success. $3 518195
650 4 $a Nuclear magnetic resonance--NMR. $3 3560258
650 4 $a Optimization. $3 891104
650 4 $a Crystallization. $3 608799
650 4 $a Amino acids. $3 558768
650 4 $a Scientific imaging. $3 3560377
650 4 $a Chromatography. $3 1073639
650 4 $a Morphogenesis. $3 810791
650 4 $a Protein expression. $3 3554057
650 4 $a Nitrogen. $3 1314426
650 4 $a Crystal structure. $3 3561040
650 4 $a Polymers. $3 535398
650 4 $a Transmission electron microscopy. $3 567074
650 4 $a Mass spectrometry. $3 551172
650 4 $a Glycerol. $3 3685180
650 4 $a Cloning. $3 571606
650 4 $a Flexibility. $3 3560705
650 4 $a Design. $3 518875
650 4 $a Molecular weight. $3 3683783
650 4 $a Crystal lattices. $3 556010
650 4 $a Crystallography. $3 518393
650 4 $a Analytical chemistry. $3 3168300
650 4 $a Chemistry. $3 516420
650 4 $a Developmental biology. $3 592588
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650 4 $a Polymer chemistry. $3 3173488
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