Identification of protein-bound riboflavin in rat hepatocyte plasma membrane as a source of autofluorescence.
The presence of flavin compound(s) giving a yellowish-green autofluorescence in rat hepatocyte plasma membrane has recently been reported (Nokubo, M. et al. (1988) Biochim. Biophys. Acta 939, 441-448). The fluorophore can quantitatively be extracted with water at 80 degrees C from isolated plasma membranes. Gel filtration of the extract eluted with water showed two peaks, the fluorescence of which closely resembled that of riboflavin. The major peak comigrated with proteins and the minor one displayed a position identical to authentic riboflavin. When the components of the major peak were rechromatographed after acetic acid treatment and eluted with 20 mM of acetic acid, the fluorescent compound separated from the proteins and eluted at the same position as riboflavin. In paper chromatography and HPLC, the behavior of the fluorescent compound (separated by acid treatment from the proteins) was identical to that of riboflavin. SDS gel filtration of subcellular fractions of rat liver revealed that riboflavin was the dominant flavin, whereas FAD and FMN were not detectable in the plasma membrane. Microsomes and mitochondria contain predominantly FAD and FMN, and only minor quantities of riboflavin. The presence of riboflavin in the plasma membrane is a novel finding, the functional significance of which is still unclear; however, a hypothesis can be forwarded on the basis of the ability of flavins to generate superoxide anion radicals during their autoxidation.
Nokubo
M
M
First Laboratory of Clinical Physiology, Tokyo Metropolitan Institute of Gerontology, Japan.
Ohta
M
M
Kitani
K
K
Nagy
I
I
D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D066298:In Vitro Techniques; D008099:Liver / Q000502:physiology* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000502:physiology*; D008856:Microscopy, Fluorescence; D051381:Rats; D012256:Riboflavin / Q000502:physiology*; D013347:Subcellular Fractions / Q000032:analysis
D008565:Membrane Proteins; D012256:Riboflavin