X-ray scattering with momentum transfer in the plane of membrane. Application to gramicidin organization.
We demonstrate a technique for measuring x-ray (or neutron) scattering with the momentum transfer confined in the plane of membrane, for the purpose of studying lateral organization of proteins and peptides in membrane. Unlike freeze-fracture electron microscopy or atomic force microscopy which requires the membrane to be frozen or fixed, in-plane x-ray scattering can be performed with the membrane maintained in the liquid crystalline state. As an example, the controversial question of whether gramicidin forms aggregates in membrane was investigated. We used dilauroylphosphatidylcholine (DLPC) bilayers containing gramicidin in the molar ratio of 10:1. Very clear scattering curves reflecting gramicidin channel-channel correlation were obtained, even for the sample containing no heavy atoms. Thallium ions bound to gramicidin channels merely increase the magnitude of the scattering curve. Analysis of the data shows that the channels were randomly distributed in the membrane, similar to a computer simulation of freely moving disks in a plane. We suggest that oriented proteins may provide substantial x-ray contrast against the lipid background without requiring heavy-atom labeling. This should open up many possible new experiments.
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Physics Department, Rice University, Houston, Texas 77251-1892.
Ludtke
S J
SJ
Wu
Y
Y
Huang
H W
HW
D055592:Biophysical Phenomena; D001703:Biophysics; D003198:Computer Simulation; D006096:Gramicidin / Q000737:chemistry*; D007473:Ion Channels / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D008567:Membranes, Artificial; D010713:Phosphatidylcholines / Q000737:chemistry; D012542:Scattering, Radiation
D007473:Ion Channels; D008051:Lipid Bilayers; D008565:Membrane Proteins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D006096:Gramicidin; C015153:1,2-dilauroylphosphatidylcholine