Proteins are dynamic molecular machines having structural flexibility that allows conformational changes(1,2). Current methods for the determination of protein flexibility rely mainly on the measurement of thermal fluctuations and disorder in protein conformations(3-5) and tend to be experimentally challenging. Moreover, they reflect atomic fluctuations on pico-second timescales, whereas the large conformational changes in proteins typically happen on micro- to millisecond timescales(6,7). Here, we directly determine the flexibility of bacteriorhodopsin-a protein that uses the energy in light to move protons across cell membranes-at the microsecond timescale by monitoring force-induced deformations across the protein structure with a technique based on atomic force microscopy. In contrast to existing methods, the deformations we measure involve a collective response of protein residues and operate under physiologically relevant conditions with native proteins.