The Golden Mile fault zone is a key controlling structure to the estimated 75 Moz gold endowment of the Kalgoorlie gold camp in the Yilgarn craton of Western Australia. The earliest structures in the fault are F1 folds that developed during D1 recumbent-fold and thrust deformation (<2685 ± 4 Ma). These F1 folds are overprinted by a pervasive NW- to NNW-striking S2 cleavage related to sinistral shearing beginning with 2680 ± 3 Ma D2a sinistral strike-slip and culminating with ca. 2660 Ma D2c sinistral-reverse movement. The majority of deformation in the fault zone correlates to ca. 2675 Ma D2b deformation, which is characterized by sinistral-normal kinematic indicators. Late, ca. 2650–2640 Ma D3 dextral-reverse kinematic indicators overprint the earlier D2 structures. Pyrrhotite-chalcopyrite-pyrite-sphalerite-galena assemblages were emplaced throughout the D2 event within NE-trending D2a tensile fractures, NW- to NNW-striking D2b normal faults and associated breccias, and NW- to NNW-striking D2c low-angle veins, with the latter D2b and D2c structures correlating to the Fimiston and Oroya mineralization types, respectively. All D2a-, D2b-, and D2c-related sulfides in the Golden Mile fault zone show similarly restricted δ34S (~1.0–4.5‰) and elevated Δ33S (~2.0–3.0‰) values that reflect strong local sulfur contribution from shales of the Lower Black Flag Group and host-rock buffering of hydrothermal fluids related to the Fimiston and Oroya mineralization events. This host-rock buffering decreased fluid fO2, favoring the development of pyrrhotite-pyrite stable sulfide assemblages and causing respective decreases and increases in fluid Au-Te and Pb-Bi-Sb concentrations. At the camp scale, the Golden Mile fault zone exerted a primary control on the distribution of porphyry dikes and gold deposits; however, magma and hydrothermal fluid circulation was favored in adjacent, higher-order structural sites due to the fault zone’s incompetent rheology and tendency for ductile deformation and diffuse fluid flow. Other Archean examples such as Au deposits of the Larder Lake-Cadillac deformation zone in the Superior craton illustrate that this type of diffuse fluid flow in large-scale crustal fault zones can result in disseminated economic mineralization. However, this study highlights that host-rock effects on fluid chemistry in large-scale crustal fault zones exercises a strong control on a fluid’s propensity to form ore. The results of this study emphasize that both the rheology and chemistry of rocks within and adjacent to large-scale deformation zones act as important controls on the formation of gold ore in Archean terranes.