The glycosylation of proteins in fibroblasts from people with the premature 
ageing disease Hutchinson-Gilford Progeria Syndrome (progeria) was investigated. 
Protein was prepared from fibroblast cell lines established from skin biopsy 
taken from progeria patients and control donors. Glycoproteins were labelled by 
the covalent attachment of the steroid hapten digoxygenin to the sugar group. 
After separation of total protein by SDS-PAGE and electroblotting onto 
Immobilon-PTM, glycoproteins were detected by enzyme immunoassay. We have 
observed a glycoprotein of M(r) 200 kDa which is consistently present in protein 
preparations from progeria fibroblasts and which is absent, or markedly reduced, 
in preparations from control fibroblasts. This suggests that it may be useful as 
a marker for progeria. Similar analysis of progeria lymphoblast and control 
lymphoblast cultures did not show this altered pattern of glycosylated proteins, 
indicating that it may be cell-type specific. Glycoproteins were also detected 
by labelling fibroblasts in vitro with D-[6-3H]glucosamine hydrochloride 
followed by SDS-PAGE of isolated protein and subsequent fluorography. Profiles 
of glycoproteins from progeria and control fibroblasts were consistent with 
those obtained from labelling of carbohydrate groups with digoxygenin. Protease 
digestion of cell protein verified that the band at M(r) 200 kDa contains a 
protein core. Characteristic features of progeria primarily involve the 
connective tissue and include wrinkled and loose skin, loss of soft tissue, thin 
limbs and stiff joints. Death of progeria patients is usually a result of 
cardiovascular abnormalities. The most consistent manifestations thus involve 
the connective tissue. The glycoprotein of M(r) 200 kDa which we have observed 
in progeria fibroblasts in vitro could reflect a perturbation in glycosylation 
which may underly the connective tissue defects seen in progeria.