Nerve growth factor-induced changes in the structure of sulfated proteoglycans in PC12 pheochromocytoma cells.
Structural changes in proteoglycans (PGs) were examined during the neuritogenesis of PC12 cells induced by nerve growth factor (NGF). (1) A heparan sulfate (HS) PG and a chondroitin sulfate (CS) PG were synthesized by PC12 cells, irrespective of the presence of NGF or the duration of culture. PGs released from PC12 cells into the culture medium were mostly CSPGs. (2) In the absence of NGF, the apparent molecular mass of HSPG prepared from PC12 cells after 3 days of culture was in the range of 90-190 kDa for the intact form (Kav = 0.38 on Sepharose CL-6B), 12 kDa for HS, and 61 kDa for the core protein. In the presence of NGF, these values were 90-190 kDa, 10 kDa, and 51 kDa and 61 kDa, respectively. The intact forms of cell-associated CSPG had apparent molecular mass ranges of 120-150 kDa and 120-190 kDa (Kav = 0.38 and 0.34), with CSs of 15 kDa and 20 kDa in the presence and absence of NGF, respectively. The apparent molecular mass of the core protein of cell-associated CSPG was 92 kDa, irrespective of the presence of NGF. The molecular sizes of cell-associated PGs and their glycosaminoglycans remained unchanged during culture. (3) CSPGs released by PC12 cells into the culture medium were separated into two peaks (I and II) by column chromatography on DEAE-cellulose. The peak II fraction prepared from the medium with NGF after 3 days of culture consisted of CSPG with Kav = 0.22 on Sephacryl S-300 [40-84 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)].(ABSTRACT TRUNCATED AT 250 WORDS)
Katoh-Semba
R
R
Department of Perinatology, Aichi Prefecture Colony, Japan.
Oohira
A
A
Kashiwamata
S
S
D000818:Animals; D002845:Chromatography; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000494:pharmacology*; D016716:PC12 Cells / Q000378:metabolism*; D011509:Proteoglycans / Q000302:isolation & purification / Q000378:metabolism*
D003470:Culture Media; D009414:Nerve Growth Factors; D011509:Proteoglycans