Domain structure of phytochrome from Avena sativa visualized by electron microscopy

Highly purified phytochrome from Avena sativa was visualized by electron microscopy after negative staining with uranyl acetate and after rotary shadowing with platinum. The particle shape was variable in both types of specimens, but tripartite structures resembling a 'Y' were consistently observed. The tripartite substructure is composed of three globular domains each having a diameter of 7 to 8 nm and equally spaced in an equilateral triangle. The dimensions of the tripartite particle measured 15 nm between the centers of any two of the three particles. When phytochrome was digested with trypsin in a manner which releases the amino-terminal globular domain from the polypeptide, the tripartite structure was lost and only small globular particles were seen. We propose that the outer particles of this tripartite structure are the amino-terminal domains of the phytochrome dimer, and the central particle comprises the carboxyl domains of the two subunits.