Revisiting the human polypeptide GalNAc-T1 and T13 paralogs

Glycobiology. 2017 Jan;27(2):140-153. doi: 10.1093/glycob/cww111. Epub 2016 Nov 22.

Abstract

Polypeptide GalNAc-transferases (GalNAc-Ts) constitute a family of 20 human glycosyltransferases (comprising 9 subfamilies), which initiate mucin-type O-glycosylation. The O-glycoproteome is thought to be differentially regulated via the different substrate specificities and expression patterns of each GalNAc-T isoforms. Here, we present a comprehensive in vitro analysis of the peptide substrate specificity of GalNAc-T13, showing that it essentially overlaps with the ubiquitous expressed GalNAc-T1 isoform found in the same subfamily as T13. We have also identified and partially characterized nine splice variants of GalNAc-T13, which add further complexity to the GalNAc-T family. Two variants with changes in their lectin domains were characterized by in vitro glycosylation assays, and one (Δ39Ex9) was inactive while the second one (Ex10b) had essentially unaltered activity. We used reverse transcription-polymerase chain reaction analysis of human neuroblastoma cell lines, normal brain and a small panel of neuroblastoma tumors to demonstrate that several splice variants (Ex10b, ΔEx9, ΔEx2-7 and ΔEx6/8-39bpEx9) were highly expressed in tumor cell lines compared with normal brain, although the functional implications remain to be unveiled. In summary, the GalNAc-T13 isoform is predicted to function similarly to GalNAc-T1 against peptide substrates in vivo, in contrast to a prior report, but is unique by being selectively expressed in the brain.

Keywords: GALNT/GALNT13/splicing/MALDI-TOF/neuroblastoma/cancer.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Brain / metabolism
  • Gene Expression Regulation
  • Glycopeptides / genetics*
  • Glycopeptides / metabolism
  • Glycosylation
  • Humans
  • Lectins / genetics
  • Lectins / metabolism
  • N-Acetylgalactosaminyltransferases / genetics*
  • N-Acetylgalactosaminyltransferases / metabolism
  • Peptides / genetics*
  • Peptides / metabolism
  • Protein Isoforms / genetics*
  • Protein Isoforms / metabolism
  • Substrate Specificity

Substances

  • Glycopeptides
  • Lectins
  • Peptides
  • Protein Isoforms
  • N-Acetylgalactosaminyltransferases
  • polypeptide N-acetylgalactosaminyltransferase