Enzymes for O-glycans

Enzymes for O-glycans

OpeRATOR™

OpeRATOR is an O-protease digesting O-glycosylated proteins N-terminally of the S/T glycosylation site. The presence of O-glycans is required for OpeRATOR to exert its enzymatic activity.

OpeRATOR is supplied as a lyophilized powder with no preservatives added, and is available in 2000 unit vials for digestion of up to 2 mg O-glycosylated protein. The enzyme is shipped with 2000 units of SialEXO for removal of sialic acids.

 

 

Detailed Information

Product Description

OpeRATOR is an endoprotease that catalyzes the hydrolysis of peptide bonds in native O-glycosylated proteins in direct proximity of the O-glycans. With high specificity, OpeRATOR digests proteins at the N-terminus of O-glycans at serine or threonine. O-glycans are required for OpeRATOR activity and the enzyme will not digest glycoproteins with only N-linked glycans. OpeRATOR is active on O-glycan proteins with or without sialic acids, although the activity is higher on O-glycan proteins where sialic acids have been removed. OpeRATOR can be used together with SialEXO (sialidase), which is included in the box. 

OpeRATOR is derived from Akkermansia muciniphila and expressed in E. coli. The enzyme contains a His-tag and the molecular weight is 42 kDa. SialEXO is derived from Akkermansia muciniphila and expressed in E. coli. The enzymes in the SialEXO also contain a His-tag and the molecular weights of these components are 43 kDa and 66 kDa, respectively.

Unit Definition

One unit OpeRATOR 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.

Content and Storage

The OpeRATOR box includes:

  • 1 vial of OpeRATOR enzyme (G1-OP1-020) supplied lyophilized in TBS pH 7.6, with no preservatives added.
  • 1 vial of SialEXO (G1-SM1-020) supplied lyophilized in TBS pH 7.6, with no preservatives added.

The vials in the OpeRATOR box is shipped cold and should be stored at –20 °C upon arrival. After reconstitution the enzymes are stable for 1 month at +4-8 °C.

 

 

Download Brochure

O-glycans_Produktfolder_Webb_20170601_OG1706-1

 

Applications

  • O-glycan profiling
  • O-glycan site determination
  • Middle-down approaches for O-glycosylated proteins
  • O-glycopeptide mapping

 

Key Characteristics

  • O-glycan specific endoprotease
  • Digests N-terminally of O-glycan-occupied serine or threonine
  • Requires O-glycan for activity
  • No activity on glycoproteins with only N-glycans
  • Pretreatment with SialEXO increases OpeRATOR enzyme performance

How to order

For details on ordering please visit How to order. Click add to cart above to buy online.

Quotations

Please do not hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities or licensing opportunities.

Documents

Instructions

 

 

Product specifications

 

 

Safety Data Sheet

 

 

FAQ and Support

Yes, the enzymes work in combination and can be used to desialylate and digest an O-glycosylated protein.

Patent and Disclaimer

OpeRATOR™

All rights reserved. Aspects of OpeRATOR™ technology are encompassed by pending patent applications in the name of Genovis AB.

The trademark OpeRATOR™ is the property of Genovis AB.

For research use only. Not intended for any animal or human therapeutic or diagnostic use.

©2017 Genovis AB.