Journal: iScience

Article Title: A pain-causing and paralytic ant venom glycopeptide

doi: 10.1016/j.isci.2021.103175

Figure Lengend Snippet: Glycosylation is critical to maintaining the aqueous solubility of Mg7a (A) Enzymatic deglycosylation of Mg7a. Time-course of deglycosylation of Mg7a, as measured by relative ion count of glycoforms at 1, 2 and 4 h of treatment with α- N -acetylgalactosaminidase. At 1 h, only 2% of detected peptide was still fully glycosylated, while 91% was fully deglycosylated and 7% was partially deglycosylated (i.e., Mg7a with the loss of 2 GalNAc moieties). At 2 h, no fully glycosylated peptide was detected, and 94% was fully deglycosylated. (B) Purification of deglyco-Mg7a by HPLC on a Gemini NX-C18 column (250 × 4.6 mm; particle size, 3 μm; pore size, 110 Å; Phenomenex) using a gradient of 5 to 50% solvent B (90% MeCN and 0.05% TFA) over 45 min at a flow rate of 1 mL/min. Mass of eluting peaks was confirmed by MALDI-MS (linear mode). Theoretical monoisotopic [M+1H] 1+ of Mg7a (loss of 2 GalNAc) = m/z 6705; Mg7a (loss of 3 GalNAc) = m/z 6503. Trace from Mg7a (fully glycosylated), run under the same conditions, is shown for comparison. (C) Fibrillar precipitate formed upon reconstitution of deglycosylated Mg7a in water at a concentration of ∼100 μM. Image was acquired on a Nikon DS-Qi2 microscope; scale bar = 100 μm. (D) Activity of Mg7a and analogs on F11 cells. Data are expressed as mean ± SEM ( n = 3).

Article Snippet: To overcome this challenge, we enzymatically deglycosylated the full-length synthetic Mg7a 1 using α- N -acetylgalactosaminidase (New England BioLabs) ( A and 3B).

Techniques: Solubility, Purification, Concentration Assay, Microscopy, Activity Assay

Journal: iScience

Article Title: A pain-causing and paralytic ant venom glycopeptide

doi: 10.1016/j.isci.2021.103175

Figure Lengend Snippet:

Article Snippet: To overcome this challenge, we enzymatically deglycosylated the full-length synthetic Mg7a 1 using α- N -acetylgalactosaminidase (New England BioLabs) ( A and 3B).

Techniques: Recombinant, Software

Journal: iScience

Article Title: A pain-causing and paralytic ant venom glycopeptide

doi: 10.1016/j.isci.2021.103175

Figure Lengend Snippet:

Article Snippet: α-N-acetylgalactosaminidase , New England BioLabs , P0734S.

Techniques: Recombinant, Software