A useful radiolabeling method for detection of T cell receptor glycoproteins bearing immature and mature N-linked glycans.
The antigen receptor expressed on most T lymphocytes, the T cell receptor (TCR), is a multisubunit complex containing TCR alpha,beta and CD3-gamma,delta, epsilon and zeta chains, in the form alphabetadeltaepsilongammaepsilonzetazeta. The intracellular transport of TCR proteins is routinely measured by the extent of processing of N-linked glycan chains present on TCR alpha,beta and CD3 gamma,delta subunits, which are converted from immature (high mannose) oligosaccharides in the endoplasmic reticulum (ER) to mature (complex type) species within the Golgi complex. In the current study, a radiolabeling method is described that is useful for the study of TCR glycoproteins bearing immature and mature N-linked oligosaccharide chains. Specifically, we report the use of [3H] galactose as a radioactive tracer of TCR proteins containing glucose and galactose residues, present in immature and mature N-linked oligosaccharides, respectively. This technique is rapid, simple, and sensitive, and provides a unique approach for studying the processing and subcellular localization of T cell receptor proteins.
Kearse
K P
KP
Department of Microbiology, East Carolina University, School of Medicine, Greenville, NC, USA. kearse@brody.med.ecu.edu
D005690:Galactose / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D008715:Methionine / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis*; D011948:Receptors, Antigen, T-Cell / Q000032:analysis*; D014316:Tritium
D006023:Glycoproteins; D011134:Polysaccharides; D011948:Receptors, Antigen, T-Cell; D014316:Tritium; D008715:Methionine; D005947:Glucose; D005690:Galactose