Atomic force microscopy (AFM) was used to analyze the interactions between fibrinogen and model surfaces having different levels of water wettability. In contrast to most AFM studies, proteins were coupled to the substrate while model surface colloids were attached to the end of the AFM probe, thereby ensuring that proteins undergo only a single compression/decompression cycle. Similar values of adhesion force were observed between fibrinogen and all of the highly wettable surfaces; in the same manner, fibrinogen showed similar adhesion forces against all poorly wettable surfaces, with a step-like transition observed between the two groups. Relationships between the adhesion forces and loading rates were used to analyze the energy profiles involved in protein/surface interactions. Multiple energy barriers were found in the interaction of proteins with poorly wettable surfaces; whereas a single energy barrier was found for protein interactions with highly wettable surfaces. Contact time-dependent adhesion data were fit to an exponential model and showed that the rate constants of the protein unfolding process on highly wettable surface were smaller at low loading rates, but increased rapidly to yield values similar to those on poorly wettable surfaces at high loading rates. The activation energies of protein unfolding derived from the data offer insight into the role of surface wettability in affecting adhesion, conformational changes, and ultimately, the activity of proteins at biomaterial surfaces.