Nonfouling NTA-PEG-Based TEM Grid Coatings for Selective Capture of Histidine-Tagged Protein Targets from Cell Lysates

We report the preparation and performance of TEM grids bearing stabilized nonfouling lipid monolayer coatings. These films contain NTA capture ligands of controllable areal density at the distal end of a flexible poly(ethylene glycol) 2000 (PEG2000) spacer to avoid preferred orientation of surface-b...

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Veröffentlicht in:	Langmuir : the ACS journal of surfaces and colloids. - 1999. - 32(2016), 2 vom: 19. Jan., Seite 551-9
1. Verfasser:	Benjamin, Christopher J (VerfasserIn)
Weitere Verfasser:	Wright, Kyle J, Hyun, Seok-Hee, Krynski, Kyle, Yu, Guimei, Bajaj, Ruchika, Guo, Fei, Stauffacher, Cynthia V, Jiang, Wen, Thompson, David H
Format:	Online-Aufsatz
Sprache:	English
Veröffentlicht:	2016
Zugriff auf das übergeordnete Werk:	Langmuir : the ACS journal of surfaces and colloids
Schlagworte:	Journal Article Research Support, N.I.H., Extramural 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy-poly(ethylene glycol 2000) ATP-Binding Cassette Transporters Cell Extracts Escherichia coli Proteins His-His-His-His-His-His MPEG350 Oligopeptides Phosphatidylethanolamines mehr... Recombinant Fusion Proteins Ribosomal Proteins maltose transport system, E coli Green Fluorescent Proteins 147336-22-9 Polyethylene Glycols 3WJQ0SDW1A Histidine 4QD397987E ribosomal protein L7-L12 70815-33-7 polyethylene glycol 2000 HAF0412YIT Nitrilotriacetic Acid KA90006V9D


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100	1		\|a Benjamin, Christopher J \|e verfasserin \|4 aut
245	1	0	\|a Nonfouling NTA-PEG-Based TEM Grid Coatings for Selective Capture of Histidine-Tagged Protein Targets from Cell Lysates
264		1	\|c 2016
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500			\|a Date Completed 24.10.2016
500			\|a Date Revised 02.12.2018
500			\|a published: Print-Electronic
500			\|a Citation Status MEDLINE
520			\|a We report the preparation and performance of TEM grids bearing stabilized nonfouling lipid monolayer coatings. These films contain NTA capture ligands of controllable areal density at the distal end of a flexible poly(ethylene glycol) 2000 (PEG2000) spacer to avoid preferred orientation of surface-bound histidine-tagged (His-tag) protein targets. Langmuir-Schaefer deposition at 30 mN/m of mixed monolayers containing two novel synthetic lipids-1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[(5-amido-1-carboxypentyl)iminodiacetic acid]polyethylene glycolamide 2000) (NTA-PEG2000-DSPE) and 1,2-(tricosa-10',12'-diynoyl)-sn-glycero-3-phosphoethanolamine-N-(methoxypolyethylene glycolamide 350) (mPEG350-DTPE)-in 1:99 and 5:95 molar ratios prior to treatment with a 5 min, 254 nm light exposure was used for grid fabrication. These conditions were designed to limit nonspecific protein adsorption onto the stabilized lipid coating by favoring the formation of a mPEG350 brush layer below a flexible, mushroom conformation of NTA-PEG2000 at low surface density to enable specific immobilization and random orientation of the protein target on the EM grid. These grids were then used to capture His6-T7 bacteriophage and RplL from cell lysates, as well as purified His8-green fluorescent protein (GFP) and nanodisc solubilized maltose transporter, His6-MalFGK2. Our findings indicate that TEM grid supported, polymerized NTA lipid monolayers are capable of capturing His-tag protein targets in a manner that controls their areal densities, while efficiently blocking nonspecific adsorption and limiting film degradation, even upon prolonged detergent exposure
650		4	\|a Journal Article
650		4	\|a Research Support, N.I.H., Extramural
650		7	\|a 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy-poly(ethylene glycol 2000) \|2 NLM
650		7	\|a ATP-Binding Cassette Transporters \|2 NLM
650		7	\|a Cell Extracts \|2 NLM
650		7	\|a Escherichia coli Proteins \|2 NLM
650		7	\|a His-His-His-His-His-His \|2 NLM
650		7	\|a MPEG350 \|2 NLM
650		7	\|a Oligopeptides \|2 NLM
650		7	\|a Phosphatidylethanolamines \|2 NLM
650		7	\|a Recombinant Fusion Proteins \|2 NLM
650		7	\|a Ribosomal Proteins \|2 NLM
650		7	\|a maltose transport system, E coli \|2 NLM
650		7	\|a Green Fluorescent Proteins \|2 NLM
650		7	\|a 147336-22-9 \|2 NLM
650		7	\|a Polyethylene Glycols \|2 NLM
650		7	\|a 3WJQ0SDW1A \|2 NLM
650		7	\|a Histidine \|2 NLM
650		7	\|a 4QD397987E \|2 NLM
650		7	\|a ribosomal protein L7-L12 \|2 NLM
650		7	\|a 70815-33-7 \|2 NLM
650		7	\|a polyethylene glycol 2000 \|2 NLM
650		7	\|a HAF0412YIT \|2 NLM
650		7	\|a Nitrilotriacetic Acid \|2 NLM
650		7	\|a KA90006V9D \|2 NLM
700	1		\|a Wright, Kyle J \|e verfasserin \|4 aut
700	1		\|a Hyun, Seok-Hee \|e verfasserin \|4 aut
700	1		\|a Krynski, Kyle \|e verfasserin \|4 aut
700	1		\|a Yu, Guimei \|e verfasserin \|4 aut
700	1		\|a Bajaj, Ruchika \|e verfasserin \|4 aut
700	1		\|a Guo, Fei \|e verfasserin \|4 aut
700	1		\|a Stauffacher, Cynthia V \|e verfasserin \|4 aut
700	1		\|a Jiang, Wen \|e verfasserin \|4 aut
700	1		\|a Thompson, David H \|e verfasserin \|4 aut
773	0	8	\|i Enthalten in \|t Langmuir : the ACS journal of surfaces and colloids \|d 1999 \|g 32(2016), 2 vom: 19. Jan., Seite 551-9 \|w (DE-627)NLM098181009 \|x 1520-5827 \|7 nnns
773	1	8	\|g volume:32 \|g year:2016 \|g number:2 \|g day:19 \|g month:01 \|g pages:551-9
856	4	0	\|u http://dx.doi.org/10.1021/acs.langmuir.5b03445 \|3 Volltext
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