The electronic and chemical structure of the interface between the amino acid L-cysteine and Au was determined by photoemission spectroscopy (PES). L-cysteine was deposited by repeatedly dipping Au substrates into solutions of L-cysteine in methanol with various concentrations. To enable repeat deposition without significant contamination, the dipping procedure was performed in a glovebox directly connected to the ultrahigh vacuum (UHV) chamber in a N2 atmosphere. X-ray photoemission spectroscopy (XPS) measurements between deposition steps allowed to characterize the chemical interaction at the interface to be characterized. Ultraviolet photoemission spectroscopy (UPS) measurements yielded the orbital line-up at the interface as well as the highest occupied molecular orbital (HOMO) structure of L-cysteine. The charge injection barrier between the L-cysteine HOMO and the Au Fermi level was found to be 3.0 eV. The interface dipole between the Au substrate and the L-cysteine overlayer was determined to be 1.03 eV. The results also indicate the formation of an interface state approximately 1.5 eV above the HOMO of the L-cysteine.