Differences in the glycosylation of rat submandibular kallikreins.
The glycosylations of five different rat submandibular kallikreins, rK1, rK2, rK7, rK9 and rK10, vacuum-blotted onto nitrocellulose membranes, have been studied by means of labelled lectins using enhanced chemiluminescence detection. The results demonstrated the individual submandibular kallikreins are not heavily glycosylated in rats, but consistently show different patterns of glycosylation. Following digestion of slot-blotted enzymes with peptide-N-glycosidase F (PNGase): binding by lectin from Lens culinaris (alpha Man-directed) was abolished, whilst that of lectin from Maclura pomifera (Gal beta 1,3GalNAc-directed) persisted (but could be abolished by periodate oxidation and endo-alpha-N-acetylgalactosaminidase digestion), revealing that there are O- as well as N-linked sugar chains on the kallikreins; a novel observation for this family of enzymes. The presence of GalNAc in addition to GlcNAc, Fuc, Gal, and Man, in sugar chains of rK1 was confirmed by high pH anion exchange chromatography following acid hydrolysis. Different intensities of binding by lectin from Limax flavus (NeuNAc-directed) suggest that sialylation of individual kallikreins differs, whilst sialidase and PNGase digestions suggest that sialic acid is the terminal residue of some N-linked but not O-linked structures.
Zhang
X S
XS
Department of Oral Pathology, King's College School of Medicine and Dentistry, London, UK.
Proctor
G B
GB
Garrett
J R
JR
Shori
D K
DK
Carpenter
G H
GH
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002240:Carbohydrate Sequence; D006031:Glycosylation; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007610:Kallikreins / Q000737:chemistry* / Q000378:metabolism; D037102:Lectins; D008297:Male; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009005:Monosaccharides / Q000032:analysis; D009842:Oligopeptides / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D013363:Submandibular Gland / Q000201:enzymology*; D013379:Substrate Specificity
D007527:Isoenzymes; D037102:Lectins; D009005:Monosaccharides; D009842:Oligopeptides; D007610:Kallikreins