TY - JOUR
T1 - A method for [3H]mannose labeling of Asn-linked oligosaccharides on recombinant glycoproteins synthesized in Xenopus oocytes
AU - Cantor, Alan B.
AU - Kornfeld, Stuart
N1 - Funding Information:
structural analysis, and Debbie Sinak for assistance with preparation of this manuscript. A.B.C. was supported by a National Institutes of Health Research Award GM-07200.Medical Scientist from the National Institutes of General Medical Sciences. This work was supported in part by United States Public Health Service Grant CA 08759.
PY - 1992/9
Y1 - 1992/9
N2 - We have developed an efficient method for labeling the Asn-linked oligosaccharides of recombinant glycoproteins synthesized in Xenopus laevis oocytes. By coinjecting GDP-[3,4-3H]mannose with mRNA for human cathepsin D, it was possible to incorporate as much as 1800 cpm per oocyte into each of the two Asn-linked oligosaccharides of this glycoprotein. Overall, about 50% of the microinjected GDP-[3,4-3H]mannose was incorporated into Asn-linked oligosaccharides, a 10-fold greater value than that obtained when [2-3H]mannose was microinjected. Less than 10% of the injected GDP-[3,4-3H]mannose was metabolized to water or converted to amino acids. This technique should facilitate studies of Asn-linked oligosaccharide biosynthesis, processing, and structure in recombinant proteins synthesized in Xenopus oocytes.
AB - We have developed an efficient method for labeling the Asn-linked oligosaccharides of recombinant glycoproteins synthesized in Xenopus laevis oocytes. By coinjecting GDP-[3,4-3H]mannose with mRNA for human cathepsin D, it was possible to incorporate as much as 1800 cpm per oocyte into each of the two Asn-linked oligosaccharides of this glycoprotein. Overall, about 50% of the microinjected GDP-[3,4-3H]mannose was incorporated into Asn-linked oligosaccharides, a 10-fold greater value than that obtained when [2-3H]mannose was microinjected. Less than 10% of the injected GDP-[3,4-3H]mannose was metabolized to water or converted to amino acids. This technique should facilitate studies of Asn-linked oligosaccharide biosynthesis, processing, and structure in recombinant proteins synthesized in Xenopus oocytes.
UR - http://www.scopus.com/inward/record.url?scp=0026616764&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(92)90427-9
DO - 10.1016/0003-2697(92)90427-9
M3 - Article
C2 - 1443567
AN - SCOPUS:0026616764
SN - 0003-2697
VL - 205
SP - 220
EP - 226
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -