Melbourne, with its relatively high proportion of Scots squatters and businessmen, had prominent churches associated with three major Presbyterian denominations in the later 1840s: the Church of Scotland, the Scottish Free Church, and the United Presbyterians. A Church of Scotland congregation under the Rev. James Forbes received a land grant in Collins Street (the Scots' Church site) in 1839. In 1848, after the Scottish Disruption, Forbes left this church to found the Knox Church congregation in Swanston Street in connection with the Free Church of Australia Felix. He placed a high value on education, founding the Scots School in 1838, the John Knox School in 1848 and the Melbourne Academy (later Scotch College, Eastern Hill) in 1851. Melburnians associated with Scottish voluntarism could join a local United Presbyterian Church congregation under the Rev. A.M. Ramsay, also in Collins Street, from 1847. Catering for members of such Scottish denominations as the United Secession Church and the Relief Church (Scottish equivalents of English 'dissenters'), the United Presbyterian Church had been founded in Sydney by the Rev. J.D. Lang.
In 1859 the three major local Presbyterian denominations united to form the Presbyterian Church of Victoria, which operated out of various Collins Street assembly halls. A small minority of Ulster Presbyterians participated in this predominantly Scottish denominational life. In addition to ministering to rural congregations, Ulster Presbyterians have played major educational roles in Melbourne: John McFarland was the founding Master of Ormond College, and Chancellor of the University of Melbourne; John L. Rentoul was first Principal of Ormond College Theological Hall; Sir Samuel Wilson from County Antrim was the patron of the university's Wilson Hall, and more recently John Davis McCaughey was master of Ormond College and later governor of Victoria.
In the late 19th century Melbourne's large and fashionable Presbyterian congregations were to be found at Chalmers Church on the Eastern Hill under Dr Adam Cairns, and at Scots Church in Collins Street under the Rev. Peter Menzies and Rev. Charles Strong. Chalmers Church under Cairns was orthodox and staid, but the Scots Church congregation under Menzies and Strong gained a reputation as a progressive and theologically liberal Christian body. The future Dame Nellie Melba (whose father David Mitchell built the church) sang in the choir, and Alexander Morrison of Scotch College was a leading elder. The public furore over heresy charges instituted against Strong caused serious unrest and divisions within Melbourne Presbyterianism of the 1880s. Faced with a religious backlash against theological 'progressives', Strong and his supporters defected to found the Australian Church.
Among suburban congregations, Toorak Presbyterian Church was especially influential because of its associations with squatters and leading politicians. Francis Ormond MLC - founder of Ormond College, the Working Man's College (RMIT) and the Ormond Chair of Music at the University of Melbourne - attended Toorak Presbyterian services, as did influential merchant and squatter James Balfour MLC, who, with fellow Presbyterians John Joseph and Matthew Davies, played an inglorious role in the land speculation that led to the disastrous Melbourne bank crash in 1893.
Presbyterianism lost status with the decline in power and influence of Scottish pastoral families in the 20th century and a more general decline in Protestant religion, but Scots Church and the adjacent assembly hall remained focal points of Presbyterian life. Such major institutions as Scotch College, the Presbyterian Ladies' College, the Geelong College and Ormond College in the University of Melbourne continued to be associated with prominent Presbyterian families of largely Scottish derivation well into the 20th century. However, Presbyterianism has been more influential in rural Victoria than in Melbourne. Postwar immigration further diluted the Scottish Presbyterian tradition, with a few Dutch immigrants participating in previously Anglo-Celtic Presbyterian congregations. When a majority of Victoria's Presbyterians entered the newly formed Uniting Church in the 1970s, the Scots' Church congregation remained connected to the continuing Presbyterian body and is currently the city's main centre of Presbyterianism.