(A) Docking pose of AM6538 (magenta sticks) and predicted hydrogen bond (orange dashed lines) and π-π interaction (blue dashed lines) in comparison with AM6538 in crystal structure (green sticks). The theoretical m/z for the monoisotopic peak of AM6538 is 542.1356.Ĥ: Figure S4 MD Simulations of the Predicted Antagonists and Mutagenesis Validation of the Docking Predictions. (F) The high-resolution mass spectrometry spectra of the pure standard AM6538 (top) and the ligand dissociated from CB 1 complex (bottom). (E) Crystal images for CB 1 in complex with AM6538 (scale bar is 90 mm). (D) Superposition of AM6538 in crystal structure (green) and local minimum calculated by quantum mechanics (magenta). The receptor is shown in gray cartoon and gray surface, AM6538 is shown in cyan carbons. (C) The 2,4-dichorophenyl ring of AM6538 fits well into the shape of the side-pocket. (B) Crystal packing of the CB 1-AM6538 complex, N-terminus involved in the crystal packing. AM6538 is shown in green stick representation and FMN (the substrate of Flavodoxin) is shown in magenta stick representation. CB 1 and Flavodoxin are colored in orange and slate, respectively. (A) Overall structure of CB 1-AM6538 complex. CB 1-1: Coommassie-stained PAGE of sample purified by cobalt immobilized-metal affinity chromatography (TALON-IMAC) CB 1-2: Coommassie-stained PAGE of sample after TEV protease digestion.ģ: Figure S3 Fusion Protein Engineering and Crystal Packing of CB 1-AM6538 Complex. (D) Coommassie-stained polyacrylamide gel electrophoresis (PAGE) of the purified crystallization-grade CB 1. The Tm value of the CB 1 mutant containing all four mutations (orange trace) is higher than other mutants, indicating that this mutation combination improves protein thermostability. (C) CPM ramping assay of CB 1 mutants testing the effects of mutations and mutation combinations on protein thermostability. (B) The normalized data show that the combination (orange trace) improves protein homogeneity. (A) The raw data show that the CB 1 mutant containing all four mutations (orange trace) has the highest protein yield. (A, B) Analytical size exclusion chromatography (aSEC) of CB 1 mutants testing the effects of mutations and mutation combination on protein homogeneity. watch people on the flight deck suffer from behind the glass of the porthole in the A/C.2: Figure S2 Stability Assay of CB 1 Mutants. truth is you should pick it because you're never gunna get paid extra for doing extra work, so why not choose the kush job. most of the AGs fuck each other.literally. never do anything truly satisfying that makes you kick back and say "phew, a hard days work, i deserve some rest." lotta geeks and freaks too. make believe building power points is important.
maybe italy,hawaii,possibly germany, spain. people advance without ever really needing to try hard or learn much so the 5 year first class really doesn't know jack shit cause well. it's a community of people who barely work hard, but get advanced quickly, so you end up with this shitty, self entitled/lazy atmosphere of spoiled people.
it's one of the easiest jobs they'll ever offer, i'd only wager this against MC. 2nd was 80% last time, it'll prob shoot back up if it doesn't im not worried. great advancement, i just write my name on the page.
0 kommentar(er)
