Proline incorporation by cultured skin fibroblasts from patients with duchenne muscular dystrophy.
The incorporation of proline by cultured skin fibroblasts derived from normal individuals and patients with Duchenne muscular dystrophy (DMD) was analyzed by SDS-gel electrophoresis combined with a double-labeling procedure [Pediatr. Res., 12: 887 (1978)]. DMD fibroblasts showed increased proline incorporation into protein of approximately 130 000 daltons which could be degraded partially by collagenase. This difference was observed only at 7 days after seeding, and may be due to slight differences in growth rate as comparison of cells harvested at 2 and 7 days indicated that increased proline incorporation into high molecular weight protein was associated with day 7 cells, whether normal or DMD. During 14 days in culture normal and DMD strains did not differ in protein content, doubling time or incorporation of [3H]leucine and [14C]proline into cellular protein, although the ratio of [14C]proline to [3H]leucine incorporated was greater for DMD fibroblasts at 7 days. Incorporation of [14C]proline and [3H]leucine into extracellular proteins was greater in DMD fibroblast cultures. These subtle differences support the hypothesis that the DMD gene is expressed in fibroblasts.
D002454:Cell Differentiation; D003470:Culture Media; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007930:Leucine / Q000378:metabolism; D008970:Molecular Weight; D009136:Muscular Dystrophies / Q000378:metabolism*; D011392:Proline / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000378:metabolism*
D003470:Culture Media; D011506:Proteins; D011392:Proline; D007930:Leucine