OpeRATOR® (O-glycan specific Protease)
O-glycan Ser/Thr 잔기의 N-terminal에서 가수분해하는 단백질 분해 효소로,
O-glycan Profiling, Site Determination 등에 활용 가능
Mucin type O-glycosylated protein 의 peptide backbone 을 가수분해 합니다.
N-Glycosylated protein 에는 반응하지 않습니다.
Figure 1. Specificity for O-glycosylated proteins. The O-protease digests only O-glycosylated proteins. No activity was observed on a nonglycosylated protein, or on a protein with only N-glycans. The proteins were pre-treated with sialidases and incubated o/n with the O-protease.
Sialic acid 제거를 위해, SialEXO (Salidase) 2000 units 이 함께 동봉되어 있습니다.
(Sialic acid 를 제거하면 OpeRATOR의 Activity 가 더 높습니다)
Figure 2. OpeRATOR is active on sialylated O-glycoproteins, but the activity is higher when the sialic acids are removed using SialEXO. The native O-glycosylated TNFa receptor (TNFaR) was incubated with OpeRATOR with or without the addition of SialEXO overnight, and the reactions were separated on SDS-PAGE.
2 hr ~ overnight (16-18 hr) Reaction
Applications:
– Analysis of O-linked glycosylation
– O-glycan profiling
– O-glycan site determination
– Middle-down approaches for O-glycosylated proteins
– O-glycopeptide mapping
Unit Definition
One unit digests ≥ 90% of 1 µg glycoprotein (TNFαR) when incubated together with one unit of SialEXO in 20 mM Tris pH 6.8 at 37 °C for 2 h.
Contents
The OpeRATOR box includes:
– 1 vial of OpeRATOR enzyme (G1-OP1-020) supplied lyophilized in TBS pH 7.6
– 1 vial of SialEXO (G1-SM1-020) supplied lyophilized in TBS pH 7.6
Storage
The box is shipped at ambient temperature, and should be stored at -20°C upon arrival.
After reconstitution the enzymes are stable for 1 month at +4-8 °C.
| Part number | Item name | Digestion | Description |
| GVG2-OP1-020 | OpeRATOR® 2000 units | 2mg | Lyophilized |
| GVG3-OC6-002 | GlycOCATCH, 200 units | 200 ug | Microspin column format |