Description

PNGaseF,peptide-N-glycosidaseF,peptide-N⁴-(N-acetyl-β-glucosaminyl)asparagineamidase. ReleasesintactN-glycansbycleaving betweentheinnermostGlcNAcandAsn. 

Source:
RecombinantgenefromElizabethkingiameningosepticum,expressedinE.coli.Thesourceorganismwaspreviouslyknownas Chryseobacterium[Flavobacterium]meningosepticum[1].

Specificity: 
CleavesallN-linkedcomplex,hybridorhighmannoseoligosaccharides,unlessα(1-3)corefucosylated,asinplantglycansandsomeinsectglycans.Asparaginemustbepeptide-bondedatbothtermini.Phosphate,sulfateandsialicacidgroupsattachedtotheoligosaccharidedonotaffectcleavage.EndoFfree. 

Figure1:CleavagespecificityforGKE-5006N-Glycanase(PNGaseF).
R₁:N&CsubstitutionotherthanH
R₂: maybeHortherestofanoligosaccharide
R₃: maybeHor α(1-6)-linkedfucose

Applications:

• ReleaseofintactN-linkedglycansfromglycopeptidesandglycoproteins
• Structure-functionstudiesofN-glycosylatedglycoproteins
• Preparationofdeglycosylatedproteinsformolecularweightestimationorcrystallographystudies

Suppliedataconcentrationof ≥2.5U/ml.Foramoreconcentratedformulationof ≥10 U/ml,whichmaybeusefulfornativedigestions,seeGKE-5010BorGKE-5020B(EDTA-free).

"D"packsizedoesnotshipwithreactionbuffers thatshipwith"A" (100mU) and"B" (200mU)packsizes.Pleasecontactusifyourequirebuffers.

Formulation:
20mMTris-HCl,50mMNaCl,1mMEDTA(pH7.5)

ReactionBuffer:
5Xconcentratedbuffer,whendilutedgives20mMsodiumphosphatepH7.5,containing0.02%sodiumazide.

MW:
~35,000daltons

pHoptimum/range:
8.6/7.5-9.5

UnitDefinition:
OneunitisdefinedastheamountofenzymerequiredtocatalyzethereleaseofN-linkedoligosaccharidesfrom1μmoleofdenaturedribonucleaseBperminuteat37°C,pH7.5.

NotformulatedforFACE®analysis,useGK50050instead.

Size:1Uin400µl

Concentration: ≥2.5U/ml

ProductCode:GKE-5006D

1.KimKK,KimMK,LimJH,ParkHY,LeeST. TransferofChryseobacteriummeningosepticumandChryseobacteriummiricolatoElizabethkingiagen.nov.asElizabethkingiameningosepticacomb.nov.andElizabethkingiamiricolacomb.nov. IntJSystEvolMicrobiol.2005May;55(Pt3):1287-93.