Characterization and localization of profilin in pollen grains and tubes of Lilium longiflorum.
Pollen tubes show a rapid and dramatically polarized growth in which the actin cytoskeleton appears to play a central role. In order to understand the regulation of actin we characterized its associated protein, profilin, in pollen tubes of Lilium longiflorum. By using purified polyclonal antibodies prepared against bean root profilin [Vidali et al., 1995: Plant Physiol. 108:115-123] we detected in pollen grains and tubes two profilin polypeptides with molecular masses of 14.4 and 13.4 KDa, and an identical isoelectric point of 5.05. Profilin comprises approximately 0.47% of the total grain protein, with actin being approximately 1.4%. We were unable to detect a statistically significant profilin increase after germination, while the actin increased approximately 68%. We also spatially localized the distribution of profilin using immunocytochemistry of fixed cells at both the light and electron microscope level, and by fluorescent analog cytochemistry on live cells. The results show that profilin is evenly distributed throughout the cytoplasm and does not specifically associate with any cellular structure.
Vidali
L
L
Biology Department, University of Massachusetts, Amherst 01003, USA.
Hepler
P K
PK
D000199:Actins / Q000032:analysis; D000906:Antibodies; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002850:Chromatography, Gel; D003285:Contractile Proteins / Q000032:analysis / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D008840:Microfilament Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D008961:Models, Structural; D008970:Molecular Weight; D018521:Plant Physiological Phenomena; D010940:Plant Proteins / Q000032:analysis / Q000096:biosynthesis; D011058:Pollen / Q000502:physiology / Q000648:ultrastructure; D051304:Profilins; D017433:Protein Structure, Secondary; D003313:Zea mays
D000199:Actins; D000906:Antibodies; D000911:Antibodies, Monoclonal; D003285:Contractile Proteins; D008840:Microfilament Proteins; D010940:Plant Proteins; D051304:Profilins