The shutdown on Friday came against a backdrop of spiralling violence this week, including Israeli missile attacks that have killed three civilians in Gaza following a Palestinian bombing that killed five Israelis.
A Palestinian man fatally stabbed a 20-year-old soldier in the neck on Thursday at Qalandiya checkpoint between Jerusalem and the city of Ram Allah, the gateway for thousands of Palestinians who enter the holy city from the West Bank daily.
An Israeli army spokeswoman said: "The Qalandiya checkpoint is closed ... until further notice unless humanitarian cases need to get through."
"The Qalandiya checkpoint is closed ... until further notice unless humanitarian cases need to get through"
An Israeli army spokeswoman
Army Radio quoted a senior military officer as saying the checkpoint, part of a network that Israel maintains in the West Bank, would remain closed for several days. Israeli news media said security measures were being reviewed.
Entry to Israel by West Bank Palestinians already had been heavily restricted during a five-year-old Palestinian uprising.
Prime Minister Ariel Sharon's government ordered a "general closure" of the territory after Monday's bombing at a shopping mall in the coastal town of Netanya.
Israel says such measures are taken in self-defence to prevent further attacks.
Chief Palestinian negotiator Saeb Erikat reiterated the Palestinian accusation that Israel was imposing collective punishment that leads to "more hatred and violence".
The latest bloodshed has strained an already shaky nine-month-old truce and further diminished peace hopes stirred by Israel's Gaza withdrawal in September.