Journal: PLoS Biology

Article Title: Insight into small molecule binding to the neonatal Fc receptor by X-ray crystallography and 100 kHz magic-angle-spinning NMR

doi: 10.1371/journal.pbio.2006192

Figure Lengend Snippet: The soluble extracellular domain of neonatal Fc receptor (FcRn ECD , PDB code 1EXU) is a heterodimer composed of β2m (green) and α-chain (blue) with a cavity at the interface between the two proteins. FcRn is involved in the regulation of HSA (orange) and IgG (red) levels. The binding of both HSA and IgG to FcRn is pH dependent, which provides a mechanism for protein homeostasis through endosomal trafficking. β2m, β2-microglobulin; FcRn, neonatal Fc receptor; FcRn ECD , extracellular domain of the neonatal Fc receptor; HSA, Human Serum Albumin; IgG, Immunoglobulin G; PDB, Protein Data Bank.

Article Snippet: The coding sequences of extracellular domain (ECD) (amino acids 1–297) of human FcRn α-chain and human β2m were synthesized by Entelechon (Entelechon, Regensburg, Germany).

Techniques: Binding Assay

Journal: PLoS Biology

Article Title: Insight into small molecule binding to the neonatal Fc receptor by X-ray crystallography and 100 kHz magic-angle-spinning NMR

doi: 10.1371/journal.pbio.2006192

Figure Lengend Snippet: (A) The protein crystallized as a dimer composed of two β2m (dark grey and green) and two α-chain (light grey and blue) molecules. (B) At the interface of β2m and the α-chain, UCB-FcRn-303 (grey) occupies a binding pocket with Glycine, Cysteine, hydrophobic (Leucine), charged (Histidine, Aspartate), and polar uncharged (Serine, Glutamine) residues. β2m, β2-microglobulin; FcRn, neonatal Fc receptor; FcRn ECD , extracellular domain of the neonatal Fc receptor.

Article Snippet: The coding sequences of extracellular domain (ECD) (amino acids 1–297) of human FcRn α-chain and human β2m were synthesized by Entelechon (Entelechon, Regensburg, Germany).

Techniques: Binding Assay

Journal: PLoS Biology

Article Title: Insight into small molecule binding to the neonatal Fc receptor by X-ray crystallography and 100 kHz magic-angle-spinning NMR

doi: 10.1371/journal.pbio.2006192

Figure Lengend Snippet: (A) The soluble FcRn ECD (42 kDa) was sedimented by ultracentrifugation at 100,000 x g directly into a 0.7 mm MAS NMR rotor using a home-made filling tool. (B) 2D 15 N- 1 H correlation spectrum recorded at 100 kHz MAS of fully protonated [ 13 C, 15 N]-labeled FcRn ECD . (C) Typical linewidths of 1 H (1) and 15 N (2) at full-width-half-maximum (FWHM) of a selected cross peak from the 15 N- 1 H spectrum. β2m, β2-microglobulin; FcRn, neonatal Fc receptor; FcRn ECD , extracellular domain of the neonatal Fc receptor.

Article Snippet: The coding sequences of extracellular domain (ECD) (amino acids 1–297) of human FcRn α-chain and human β2m were synthesized by Entelechon (Entelechon, Regensburg, Germany).

Techniques: Labeling

Journal: PLoS Biology

Article Title: Insight into small molecule binding to the neonatal Fc receptor by X-ray crystallography and 100 kHz magic-angle-spinning NMR

doi: 10.1371/journal.pbio.2006192

Figure Lengend Snippet: Sequential resonance assignments using the experiments (H)CANH (blue), (H)CA(CO)NH (red), and (H)CBCANH (green) recorded on fully protonated [ 13 C, 15 N]-labeled FcRn ECD at 100 kHz MAS. As an example, the sequential connections from K41 β2m to R45 β2m in β2m are indicated by dashed lines. All assigned chemical-shifts can be found in , , and in the BMRB (accession number 27437). β2m, β2-microglobulin; BMRB, Biological Magnetic Resonance Data Bank; FcRn ECD , extracellular domain of the neonatal Fc receptor; MAS, magic-angle-spinning.

Article Snippet: The coding sequences of extracellular domain (ECD) (amino acids 1–297) of human FcRn α-chain and human β2m were synthesized by Entelechon (Entelechon, Regensburg, Germany).

Techniques: Labeling

Journal: PLoS Biology

Article Title: Insight into small molecule binding to the neonatal Fc receptor by X-ray crystallography and 100 kHz magic-angle-spinning NMR

doi: 10.1371/journal.pbio.2006192

Figure Lengend Snippet: (A) CSPs in surface representation of the FcRn ECD diprotomer crystal structure in complex with UCB-FcRn-303 (red), with the same color-coding as in . (B) For orientation, the FcRn ECD crystal structure is shown in cartoon representation with β2m in green and dark grey and the α-chain molecules in blue and light grey. (C) The IgG and HSA interaction sites are depicted in purple and orange, respectively. The highlighted residues are discussed in the text. CSP, chemical-shift perturbation; FcRn, neonatal Fc receptor; FcRn ECD , extracellular domain of the neonatal Fc receptor; HSA, Human Serum Albumin; IgG, Immunoglobulin G.

Article Snippet: The coding sequences of extracellular domain (ECD) (amino acids 1–297) of human FcRn α-chain and human β2m were synthesized by Entelechon (Entelechon, Regensburg, Germany).

Techniques:

Journal: Plant Physiology

Article Title: Targeted A-to-G base editing in the organellar genomes of Arabidopsis with monomeric programmable deaminases

doi: 10.1093/plphys/kiad678

Figure Lengend Snippet: Genotyping of T 1 plants. A) Numbers of T 1 plants with A-to-G or C-to-T mutation for 6 constructs. The numbers in parentheses represent the number of T 1 plants with homoplasmic substitution. B) Schematic of the mTALEAD construct of 16S rRNA -1 and the number of T 1 plants with bases edited and their positions for the construct 16S rRNA -1 at 11 and 24 DAS. The triangles indicate the special base-editing targets predicted to confer spm r . CTP, chloroplast targeting peptide; TALE arrays: DNA-binding sequence; AD, adenine deaminase TadA8e; DddA tox E1347A, a catalytically inactive cytidine deaminase; homo, homoplasmic substitution. The number of T 1 plants with A-to-G conversion is marked in shadow, and the nonshadow number indicates C-to-T conversion. C) Base editing results in the target of 11 and 24 DAS T 1 plants of the construct atp6-2 -2.

Article Snippet: TadA8e and DddA tox E1078K coding sequences were designed to encode the same amino acid as Cho's experiment ( ) and artificially synthesized by Twist Bioscience, with restriction enzyme cutting sites bglII and pstI at the beginning and end of the sequence.

Techniques: Mutagenesis, Construct, Binding Assay, Sequencing