One-dimensional metal/semiconductor heterojunction nanomaterials have opened many new opportunities for future nanodevices because of their novel structures and unique electrical and optical properties. In this work, sulfhydryl-containing multi-walled carbon nanotube/gold nanoparticle (MWCNT/Au) heterojunctions were synthesized in high yield by a sulfhydryl- functionalized self-assembly strategy. The component, size, structure, morphology and bond mode of the MWCNT/Au heterojunctions thus prepared were investigated and demonstrated by transmission electron microscopy, scanning electron microscopy, x-ray diffraction, energy-dispersive x-ray spectroscopy, Fourier-transform infrared and UV–visible measurements. Cyclic voltammogram and electrochemical impedance spectroscopy studies indicate that the MWCNT/Au heterojunctions have a novel electron transfer property, which retards electron transfer of the horseradish peroxidase or the ferricyanide in the underlying electrodes. We believe that MWCNT/Au heterojunctions with high stability and a unique electrical property are expected to find potential applications for nanodevices.