Molecular Cloning of Two Types of GAP Complementary DNA from Human Placenta

TLDR

The ras p21 GTPase‑activating protein was purified from human placenta, its amino‑acid sequence determined, and this sequence was used to isolate and characterize two complementary‑DNA clones. One clone encodes a 116,000‑Da GAP highly homologous to bovine GAP and expressed in multiple tissues, while the other encodes a 100,400‑Da splice variant lacking the hydrophobic N‑terminus, retains activity, and is expressed mainly in placenta and cell lines but not adult tissues.

Abstract

The ras p21 GTPase-activating protein (GAP) was purified from human placental tissue. Internal amino acid sequence was obtained from this 120,000-dalton protein and, by means of this sequence, two types of complementary DNA clones were isolated and characterized. One type encoded GAP with a predicted molecular mass of 116,000 daltons and 96% identity with bovine GAP. The messenger RNA of this GAP was detected in human lung, brain, liver, leukocytes, and placenta. The second type appeared to be generated by a differential splicing mechanism and encoded a novel form of GAP with a predicted molecular mass of 100,400 daltons. This protein lacks the hydrophobic amino terminus characteristic of the larger species, but retains GAP activity. The messenger RNA of this type was abundantly expressed in placenta and in several human cell lines, but not in adult tissues.