Effect of phosphatidylcholine structure on the adenylate cyclase activity of a murine fibroblast cell line.
To determine which structural characteristics of membrane phospholipids influence adenylate cyclase activity, we measured basal and sodium fluoride-or forskolin-stimulated activity in a murine fibroblast cell line, i.e., Balb/c3T3 cells grown in media supplemented with fetal calf serum (FCS), lipid-depleted FCS (LD-FCS) or LD-FCS complexed with different phosphatidylcholine (PC) molecular species. Cells grown in the presence of LD-FCS showed a substantial decrease in their basal and NaF-stimulated adenylate cyclase activities; however, their forskolin-stimulated activity was not altered, suggesting that the enzyme's catalytic site is not affected by changes in membrane lipids. Media supplemented with different LD-FCS/PC complexes were shown to prevent the LD-FCS-mediated reduction of basal and NaF-stimulated adenylate cyclase activity to different extents. Addition of cis-9-16:1, cis-9-18:1/cis-9-18:1 or cis-9-18:1/cis-9,12-18:2 sn-glycerophosphocholine (GPC) completely restored adenylate cyclase activity, while cis-11-18:1/cis-11-18:1 GPC was not effective and only a partial recovery was observed with 16:0/16:0, 16:0/cis-9-18:1 and trans-9-18:1 GPC. Considering the structural features of these seven PC molecular species, the findings suggest that an optimal lipid environment is conferred to the enzyme by the presence of two cis double bonds, each located in delta 9 position of the PC acyl chains. The limited effect of cis-9-16:1/cis-9-18:1 GPC and cis-9-18:1/cis-9-16:1 GPC suggests that an equal length of the terminal hydrocarbon chains extending beyond the delta 9 double bonds is also important.(ABSTRACT TRUNCATED AT 250 WORDS)
Calorini
L
L
Institute of General Pathology, University of Florence, Italy.
Mugnai
G
G
Mannini
A
A
Ruggieri
S
S
D016475:3T3 Cells; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D005576:Colforsin / Q000494:pharmacology; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D015394:Molecular Structure; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D012969:Sodium Fluoride / Q000494:pharmacology
D008563:Membrane Lipids; D010713:Phosphatidylcholines; D005576:Colforsin; D012969:Sodium Fluoride; D000262:Adenylyl Cyclases