A phospholipid is the membrane-anchoring domain of a protein growth factor of molecular mass 34 kDa in placental trophoblasts.
Recently we isolated a protein growth factor of 34 kDa from trophoblastic membranes of human placenta. A fraction (approximately equal to 50%) of the membrane-associated 34-kDa protein is peripherally associated--i.e., it can be released by high salt treatment. The remainder shows the characteristics of an integral membrane protein--i.e., its release requires detergent treatment. Here we report studies on the structural basis for membrane anchorage of the protein. Phospholipase C was found to release an immunoreactive 34-kDa polypeptide from intact isolated cytotrophoblasts. Studies with isolated trophoblastic membranes showed that phospholipase C specifically released the salt-resistant fraction of the 34-kDa polypeptide. The polypeptide released by phospholipase C showed the same electrophoretic mobility in NaDodSO4/PAGE as the polypeptide prior to phospholipase C treatment. The identity of the released protein with the 34-kDa growth factor has been established by both immunologic and receptor-binding assays. Other studies show that there is biosynthetic incorporation of [3H]myristate into the 34-kDa protein. The myristate label is labile to phospholipase C treatment. These results suggest that some of the 34-kDa protein is anchored to the plasma membrane via a posttranslationally added phospholipid. This mode of anchorage has been observed for some other membrane proteins and raises interesting questions regarding the role of this novel linkage in the mitogenic function of the 34-kDa polypeptide.
Roy-Choudhury
S
S
Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia 19104.
Mishra
V S
VS
Low
M G
MG
Das
M
M
D000215:Acylation; D000818:Animals; D005260:Female; D006133:Growth Substances / Q000032:analysis*; D006801:Humans; D008970:Molecular Weight; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011817:Rabbits; D014327:Trophoblasts / Q000032:analysis*; D010738:Type C Phospholipases / Q000378:metabolism
D006133:Growth Substances; D009227:Myristic Acids; D010743:Phospholipids; D019814:Myristic Acid; D010738:Type C Phospholipases