Beginning by the Port of Vancouver terminals near the nexus of the city's historic Downtown Eastside and Gastown districts, the street runs south through Chinatown south of Hastings Street and continues past the Pacific Central railway station and the Main Street–Science World SkyTrain station.

At Terminal Avenue, it slopes up into the Mount Pleasant neighbourhood, which mixes commerce, light industry, brewing, and high-density residential areas.

At Main Street's intersection with Marine Drive in southern Vancouver, its character becomes big-box commercial, notably including a Real Canadian Superstore supermarket.

Between Marine Drive and its southern terminus, Main Street's neighbourhood is a highly miscellaneous commercial mix including office space, manufacturing, and warehousing.

The north end of Main Street is located just west of the historic site of Hastings Mill, the nucleus around which the settlement of Granville, later Vancouver, grew.