Cryo-electron microscopy of deoxy-sickle hemoglobin fibers

Deoxy-sickle hemoglobin (HbS) polymerizes in vivo into long helical fibers which fill the red cell and make it rigid. This impedes red cell passage through the capillaries and is responsible for the clinical manifestations of sickle cell disease. Images of individual and laterally associated HbS fibers were obtained by electron microscopy of frozen hydrated specimens. Each fiber possesses variable pitch, having from 6 degrees to 12 degrees rotation per unit cell. Laterally associated HbS fibers display systematic inter-fiber contacts in spite of their pitch variations, and exhibit better order than isolated fibers. This suggest that inter-fiber contacts can act to couple fibers mechanically and might therefore be a factor in rigidifying red cells in vivo. Fiber variability was attributed to local torsional variations with a standard deviation of 2.5 degrees, but which are weakly coupled over a length of 2.25 unit cells. Variable pitch produces structural changes of as large as 5 A azimuthally and 6 A axially in HbS fiber unit cells.