Porphyrins have been in the focus of research and intense studies for over a century. Heme is an iron-containing protoporphyrin that plays variety of roles by serving as a crucial prosthetic group in hemoproteins involved in various biological processes such as oxygen transport, storage, and miRNA processing as well as a signaling molecule regulating a wide array of cellular and molecular processes. Conversely, heme also exhibits the ability to temporarily associate with proteins, influencing biochemical pathways. Moreover, due to the association to a category of diseases known as porphyrias, scientists started exploring heme and its porphyrin precursors more extensively. These include proteins and enzymes in the heme biosynthetic pathway resulting in increased levels of heme precursors. Another set of heme-related disorders affecting a large cohort worldwide, is the broad category of hemolytic diseases, encompassing sickle cell disease and β-thalassemias, which involve premature rupture of RBCs leading to the release of excess amount of heme into the plasma. This available heme can bind to various proteins and regulate their functions. Nevertheless, the understanding of heme’s contribution to these processes is still growing with increased efforts to gain insight into the mechanisms that initiate the interaction between heme and target proteins and the subsequent formation of complex structures. The presence of a distinct heme-binding motif (HBM) is a crucial requirement for the formation of such intricate complexes. While there are many short signature sequences that indicate specific protein functions, there is a lack of comprehensive analysis regarding the various patterns and structural characteristics of HBMs. Thus, the present investigation focuses on the evaluation of known heme-regulated proteins in mammals, specifically examining the specific recognition and structural patterns within their HBMs. Importantly, this analysis emphasizes the significance of Cys-Pro dipeptide motifs due to their higher frequency. Through this investigation, a comprehensive understanding of the sequence and structural similarities and differences observed during transient heme binding and the subsequent regulation of the respective proteins can be obtained.