For decades it has been known that an out-of-plane ruffling distortion of heme perturbs its UV-vis absorption (Abs) spectrum, but whether increased ruffling induces a red or blue shift of the Soret band has remained a topic of debate. This debate has been resolved by the spectroscopic and computational characterization of Mycobacterium tuberculosis MhuD presented here, an enzyme that converts heme, oxygen, and reducing equivalents to nonheme iron and mycobilin. W66F and W66A MhuD have been characterized using (1)H nuclear magnetic resonance, Abs, and magnetic circular dichroism spectroscopies, and the data have been used to develop an experimentally validated theoretical model of ruffled, ferric heme. The PBE density functional theory (DFT) model that has been developed accurately reproduces the observed spectral changes from wild type enzyme, and the underlying quantum mechanical origins of these ruffling-induced changes were revealed by analyzing the PBE DFT description of the electronic structure. Small amounts of heme ruffling have no influence on the energy of the Q-band and blue-shift the Soret band due to symmetry-allowed mixing of the Fe 3dxy and porphyrin a2u orbitals. Larger amounts of ruffling red-shift both the Q and Soret bands due to disruption of π-bonding within the porphyrin ring.