But by tweaking their technique, the researchers were able to get high-resolution images of all three portions.
"The challenge was something like having a room full of people and trying to get good photos of every one of them," said Ramamoorthy, an associate professor of chemistry and Biophysics. "With one picture, you probably can't do it. But if you say, 'Everyone over age 50 stand up,' and you take one picture, and then you ask for another age group and take another picture, and so on, you have a better chance."
By spinning their samples (or aligning the molecules in the magnetic field), the researchers were able to differentiate parts of the molecule based not on age group, as in the photo analogy, but by mobility. "With the techniques we designed, we were able to observe the rigid portion separately from the highly mobile and less mobile portions," Ramamoorthy said.
In the first part of the work, published in the Journal of the American Chemical Society in May, the researchers described the membrane-spanning segment of cytochrome b5, revealing for the first time its helical shape and the way it tilts in relation to the membrane. In the new work published in BBA Biomembranes, they determined that once both molecules are bound in the membrane, cytochrome b5 modulates the motion and the structure of cytochrome P450. More work is in progress to determine the detailed high-resolution structures of these two proteins.
Ramamoorthy's team also is studying other membrane-associated proteins, a group that includes many biologically important molecules.
"These proteins are involved in all major diseases, everywhere in the body, and are therefore primary targets for pharmaceutical applications," Ramamoorthy said. "In my opinion, solving the structures of membrane proteins should be the highest priority for structural biologists in the coming years."
|Contact: Nancy Ross-Flanigan|
University of Michigan