Journal: Immunity

Article Title: Caspase-1 engages full-length gasdermin D through two distinct interfaces that mediate caspase recruitment and substrate cleavage

doi: 10.1016/j.immuni.2020.06.007

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: NDSB-201 (Non-detergent sulfobetaine) , GOLDBIO , Cat# N-201.

Techniques: Recombinant, Modification, Mutagenesis, LDH Cytotoxicity Assay, Clone Assay, Knock-Out, Software

Journal: Journal of chemical information and modeling

Article Title: Combined Computational-Biochemical Approach Offers an Accelerated Path to Membrane Protein Solubilization.

doi: 10.1021/acs.jcim.3c00917

Figure Lengend Snippet: Figure 7. FSEC analysis indicates polydispersity for hGOAT-eGFP solubilized in DDM, GDN, and LMNG. (A) Optimized conditions increase hGOAT-eGFP solubilization in GDN, LMNG, DDM, and CHAPS. (B) FSEC analysis indicates a single peak for FOS-16 solubilized hGOAT-eGFP, while hGOAT-eGFP in DDM, GDN, and LMNG exhibit multiple peaks indicating enzyme−detergent complex polydispersity. Representative chromatograms reflect solubilizations run in triplicate on different days.

Article Snippet: Detergents were ordered as part of a solubilization screening kit (Popular Detergent Kit; 850561P-1EA, Avanti Polar Lipids) or individually for Fos-Choline-16 (F316, Anatrace) and Lauryl Maltose Neopentyl Glycol (NG310, Anatrace).

Techniques:

Journal: Journal of chemical information and modeling

Article Title: Combined Computational-Biochemical Approach Offers an Accelerated Path to Membrane Protein Solubilization.

doi: 10.1021/acs.jcim.3c00917

Figure Lengend Snippet: Figure 5. hGOAT-eGFP solubilization monitored by in-gel fluorescence. (A) Buffer sample negative control and untreated hGOAT-eGFP negative control provide size standard for fluorescent hGOAT-eGFP, with FOS-16 and FOS-12 exhibiting efficient solubilization of hGOAT-eGFP as shown by majority of fluorescence in the supernatant/soluble protein fraction. (B) hGOAT-eGFP solubilization trials exhibited partial solubilization with CHAPS, DDM, and GDN, more effective solubilization with LMNG, and little to no solubilization with MEGA-9 and BOG.

Article Snippet: Detergents were ordered as part of a solubilization screening kit (Popular Detergent Kit; 850561P-1EA, Avanti Polar Lipids) or individually for Fos-Choline-16 (F316, Anatrace) and Lauryl Maltose Neopentyl Glycol (NG310, Anatrace).

Techniques: Fluorescence, Negative Control

Journal: Journal of chemical information and modeling

Article Title: Combined Computational-Biochemical Approach Offers an Accelerated Path to Membrane Protein Solubilization.

doi: 10.1021/acs.jcim.3c00917

Figure Lengend Snippet: Figure 6. MEGA-9 maintains octanoylation activity in its hGOAT-eGFP supernatant fraction. Each hGOAT-eGFP solubilization supernatant fraction was assessed for ghrelin octanoylation activity. Reaction lacking the acyl donor served as a negative control, with untreated WT hGOAT- eGFP and the pellet from the buffer-treated hGOAT-eGFP serving as positive controls. Only the supernatant fraction from the MEGA-9 solubilizations exhibited significant activity with ∼20% conversion of substrate to octanoylated product. Activity screening reactions were performed in duplicate and analyzed as described in the Methods section.

Article Snippet: Detergents were ordered as part of a solubilization screening kit (Popular Detergent Kit; 850561P-1EA, Avanti Polar Lipids) or individually for Fos-Choline-16 (F316, Anatrace) and Lauryl Maltose Neopentyl Glycol (NG310, Anatrace).

Techniques: Activity Assay, Negative Control