Early in the city's history, many foreign countries opened their embassies on 16th Street because of its proximity to the White House. Many religious denominations followed with churches, earning the street the nickname "Church Row." These include Foundry Methodist, First Baptist, the Nineteenth Street Baptist Church which was originally named the First Colored Baptist Church of Washington, D.C., St. John's, All Souls Unitarian, Universalist National Memorial Church, St. John the Baptist Russian Orthodox Cathedral, founded in 1949 and built in 1958, and Third Church of Christ, Scientist, which was designed by an associate of I.M. Pei in 1971 and demolished in 2014. Shrine of the Sacred Heart is located just off of 16th Street. After most of the embassies moved to Embassy Row and other parts of the city, the churches became more prominent in 16th Street's identity. Other notable buildings include the Scottish Rite Masons' House of the Temple, Carnegie Institution for Science, Robert Simpson Woodward House, the Warder Mansion, Carter Barron Amphitheater, the Washington, D.C. Jewish Community Center, and the Toutorsky Mansion. The AFL-CIO, American Trucking Association, National Education Association, American Chemical Society, National Geographic Society, and Benjamin Franklin University have prominent buildings on 16th Street. The National Rifle Association was until the late 1990s headquartered on the street. The northern and central portions of 16th Street — and the Crestwood neighborhood, in particular — have for a half century been the chosen neighborhood of accomplished African Americans in Washington. Known colloquially as "The Gold Coast", these sections of 16th Street are lined with early 20th-century Tudor mansions. As 16th Street continues north through the Shepherd Park neighborhood, the street passes Tudor-style house at 7700 16th Street NW, the scene of a notorious crime, and several houses of worship including the Ohev Sholom synagogue and historic Tiffereth Israel synagogue, across the street from one another, and the Washington Ethical Society. The street's proximity to Rock Creek Park and importance as a thoroughfare has made it a natural dividing boundary for Washington neighborhoods. Outside of the downtown area, no neighborhood in the city falls on both sides of 16th Street; the neighborhoods that surround it have 16th as either their eastern or their western boundary. For many years, the wide street was the de facto "boundary" between Caucasian and African-American neighborhoods of the city, especially in the tense years after the 1968 race riots. A pair of similarly named streets, 16th Street Northeast and 16th Street Southeast, are three miles away in the northeast and southeast quadrants of Washington. They are contiguous with each other and parallel to 16th Street NW. There is no 16th Street Southwest, as this space is occupied by the National Mall and the Washington Channel.
In 1920, more than 500 trees were planted along 16th Street between Alaska Avenue and Varnum Street to honor fallen soldiers from World War I. Today, the 16th Street World War I Memorial Trees and their corresponding markers have largely been lost to history.
In July 2005, just before Congress's summer recess, TexasRepublican congressman Henry Bonilla quietly introduced resolution to rename 16th Street NW "Ronald Reagan Boulevard" in honor of the former president of the United States. Mayor Anthony A. Williams objected on the grounds that the proposal changes Pierre L'Enfant's 1791 design for the city and would have cost an estimated $1 million for new signs and maps. The plan was ultimately quashed by Rep. Tom Davis, chairman of the House Government Reform Committee and a fellow Republican representing Washington's Virginia suburbs.
On June 5, 2020, DC Public Works Department painted the words "Black Lives Matter" in 35-foot yellow capital letters on 16th Street NW near the White House and Lafayette Square, with the assistance of the MuralsDC program of the D.C. Department of Public Works. The D.C. flag accompanies the text. Later that day Mayor Muriel Bowser announced that part of the street outside of the White House had been officially renamed to Black Lives Matter Plaza. On Saturday, June 6, 2020, activists altered the mural. The stars were removed from the D.C. flag, changing it to an equals sign, and the words "defund the police" were added, resulting in the mural reading "Black Lives Matter = Defund the Police".