recombinant human lamp2 c-terminal 10 his-tag (seq id 40  (R&D Systems)

Journal: Current Research in Insect Science

Article Title: Structure of an antennally-expressed carboxylesterase suggests lepidopteran odorant degrading enzymes are broadly tuned

doi: 10.1016/j.cris.2023.100062

Figure Lengend Snippet: Crystal structure of EposCCE24 (in green), superimposed with the structure of DmelEst6 (in orange, PDB code 5THM).

Article Snippet: The C-terminally His 10 -tagged EposCCE24 construct was synthesised (Genscript, Piscataway, NJ, USA), gateway cloned into the pDEST8 vector and transformed into DH10Bac cells as per the manufacturer's protocol.

Techniques:

Journal: Current Research in Insect Science

Article Title: Structure of an antennally-expressed carboxylesterase suggests lepidopteran odorant degrading enzymes are broadly tuned

doi: 10.1016/j.cris.2023.100062

Figure Lengend Snippet: Comparisons of the internal binding cavities of EposCCE24 (A) and DmelEst6 (B). Polar atoms are shown in red (oxygen) and blue (nitrogen). Key residues lining the binding cavities are shown in stick mode. The white arrows indicate the entrances of the cavities, while the white star indicates the position of the subsite in DmelEst6 (absent in EposCCE24).

Article Snippet: The C-terminally His 10 -tagged EposCCE24 construct was synthesised (Genscript, Piscataway, NJ, USA), gateway cloned into the pDEST8 vector and transformed into DH10Bac cells as per the manufacturer's protocol.

Techniques: Binding Assay

Journal: Current Research in Insect Science

Article Title: Structure of an antennally-expressed carboxylesterase suggests lepidopteran odorant degrading enzymes are broadly tuned

doi: 10.1016/j.cris.2023.100062

Figure Lengend Snippet: Docking positions of (A) Z11–14:OAc, (B) E11–14:OAc, and (C) 14:OAc inside the binding cavity of EposCCE24. Compounds are shown in stick mode, with carbon atoms in orange (Z11–14:OAc), green (E11–14:OAc) or teal (14:OAc) and oxygen atoms in red. EposCCE24 carbon atoms are shown in green, while polar atoms are shown in blue (nitrogen) and red (oxygen).

Article Snippet: The C-terminally His 10 -tagged EposCCE24 construct was synthesised (Genscript, Piscataway, NJ, USA), gateway cloned into the pDEST8 vector and transformed into DH10Bac cells as per the manufacturer's protocol.

Techniques: Binding Assay

Journal: Current Research in Insect Science

Article Title: Structure of an antennally-expressed carboxylesterase suggests lepidopteran odorant degrading enzymes are broadly tuned

doi: 10.1016/j.cris.2023.100062

Figure Lengend Snippet: Degradation of plant volatiles and lepidopteran sex pheromone compounds by recombinant EposCCE24 as determined by GC–MS analyses. A) results from screening experiments, and B) results from kinetic experiments using the behaviourally active compounds, E11–14:OAc and Z11–14:OAc. Results represent the mean (+/- SEM) result from three biological replicates. Orange bars indicate compounds known to elicit electrophysiological responses in E. postvittana antennae.

Article Snippet: The C-terminally His 10 -tagged EposCCE24 construct was synthesised (Genscript, Piscataway, NJ, USA), gateway cloned into the pDEST8 vector and transformed into DH10Bac cells as per the manufacturer's protocol.

Techniques: Recombinant, Gas Chromatography-Mass Spectrometry

Journal: Nature Communications

Article Title: Inositol pyrophosphates activate the vacuolar transport chaperone complex in yeast by disrupting a homotypic SPX domain interaction

doi: 10.1038/s41467-023-38315-w

Figure Lengend Snippet: a Binding affinities of Vtc2* to Vtc4*, SPX2 to Vtc4*, and SPX2 to TTM4, as determined by microscale thermophoresis (MST). b Binding affinities of SPX2 to Vtc4* in the presence of 2.5 mM IP 6 , 5-IP 7 , or 1,5-IP 8 determined by MST. Cartoons illustrate the constructs used in each experiment. The resulting dissociation constants are given along with a 95% confidence interval or as a lower limit. Graphs show the means and SEM; n = 3 replicates. Source data are provided as a Source Data file.

Article Snippet: The cation exchange chromatography step was skipped for SPX2∆α1 due to protein instability caused by the pH jump and it was slightly modified for SPX2∆α7, such that a different buffer B was used (20 mM MES, pH 6.0 and 20 mM NaCl). pET27b(+)-based Vtc2(1–553), Vtc2*, and Vtc4(1–487), Vtc*, as well as TTM4 of Vtc4(192–487) with a C-terminal TEV cleaving site and a His 10 -tag were ordered from GenScript .

Techniques: Binding Assay, Microscale Thermophoresis, Construct

Journal: mBio

Article Title: InvL, an Invasin-Like Adhesin, Is a Type II Secretion System Substrate Required for Acinetobacter baumannii Uropathogenesis

doi: 10.1128/mbio.00258-22

Figure Lengend Snippet: InvL has predicted structural similarity to InvA of Yersinia . (A) The crystal structure of the passenger domain of Yersinia InvA (PDB entry 1CWV ) is shown on the left, and the AlphaFold2 predicted structure of the analogous region of InvL is pictured on the right. (B) Amino acids corresponding to subdomains of the passenger domain of InvA and the predicted homologous region of InvL. (C) Graphic depiction of the structure of InvA and the predicted structure of InvL; created with BioRender.com. Red, green, and blue colors denote individual IG-like domains, and yellow denotes the C-terminal IG-like domain in intimate contact with the lectin-like domain. βB, β-barrel domain; SP, signal peptide; OM, outer membrane.

Article Snippet: To generate recombinant protein for antibody generation, full-length InvL with a C-terminal His 10 tag was expressed from pET-22b(+):: invL - his 10 in Rosetta-gami 2(DE3) cells (Novagen, Madison, WI) using ZYM-5052 autoinducible medium for 72 h at 20°C ( ).

Techniques: