Skip to Content
MilliporeSigma
  • Development of a method for the efficient release of N-glycans from glycoproteins generating native deglycosylated proteins.

Development of a method for the efficient release of N-glycans from glycoproteins generating native deglycosylated proteins.

Enzyme and microbial technology (2012-07-05)
Shengjun Wang, Chuanliang Ma, Huili Yu, Changgan Huang, Qingsheng Qi
ABSTRACT

In many cases, it is desirable to maintain the native status of the target glycoproteins when they are deglycosylated. However, most conventional deglycosylation process often causes the irreversible denaturation of the target glycoproteins. In the present study, we developed a deglycosylation method that could obtain the native deglycosylated proteins employing Png1p-ΔH1, which was confirmed to tolerate high concentration of dithiothreitol (DTT). To prove this process, ribonuclease B (RNase B) and Yeast carboxypeptidase (CPY) were employed as the targeting glycoproteins. Our results confirmed that both of them could be completely deglycosylated in the presence of high concentration DTT and could be refolded when DTT was removed. The circular dichroism spectroscopy (CD) measurement of refolded CPY and RNase B indicated that the structure of deglycosylated proteins had recovered their native status. This method offers the possibility of efficiently releasing N-linked glycans from glycoproteins and obtaining the native target proteins.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, ready-to-use solution, recombinant, expressed in E. coli
Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, lyophilized powder, recombinant, expressed in E. coli
Sigma-Aldrich
Glycopeptidase A from almonds, buffered aqueous glycerol solution, ≥0.05 unit/mL
Sigma-Aldrich
PNGase F from Elizabethkingia miricola, buffered aqueous solution
Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, BioReagent, ≥95% (SDS-PAGE), for proteomics
Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, recombinant, expressed in E. coli, set of 100 units nanomolar unit