The organizing pastor of First Presbyterian Church was John Witherspoon, grandson and namesake of the only clergyman to sign the Declaration of Independence. The elder Witherspoon was also the President of the College of New Jersey (Princeton), delegate to the Continental Congress, and first moderator of the General Assembly of the Presbyterian Church in the United States of America in 1789.

Among the original members of First Presbyterian Church were Eunice Edwards Hunt and Frances Pollock Devereux, daughter and granddaughter of the Yale educated preacher of the “Great Awakening” Jonathan Edwards, whose writings place him in the ranks of the greatest American theologians. Their nephew and cousin, respectively, was Aaron Burr, the third Vice-President of the United States. Commemorative plaques for all thirteen founding members are placed on the walls of the sanctuary.

The sanctuary was built in 1819-1821 by architect and builder Uriah Sandy in Federal style unique in North Carolina. It is listed on the National Register of Historic Places and is an American Presbyterian and Reformed Historical Site. Adjacent to the sanctuary is the Session House built in 1858. Directly behind the sanctuary facing Johnson Street is the church manse, built in 1820 and purchased by the congregation in 1842. It is the oldest manse in continuous use in North Carolina.