Vacuolar Storage Proteins Are Sorted in the cis-Cisternae of the Pea Cotyledon Golgi Apparatus

doi:10.1083/jcb.152.1.41

Published online 2 January 2001. doi:10.1083/jcb.152.1.41

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© The Rockefeller University Press, 0021-9525/2001/1/41/ $5.00
The Journal of Cell Biology, Volume 152, Number 1, January 8, 2001 41-50

Original Article

Vacuolar Storage Proteins Are Sorted in the cis-Cisternae of the Pea Cotyledon Golgi Apparatus

Stefan Hillmer^a, Ali Movafeghi^a, David G. Robinson^a, and Giselbert Hinz^a
^a Department of Structural Cell Physiology, Albrecht-von-Haller Institute for Plant Sciences, University of Göttingen, 37073 Göttingen, Germany

Correspondence to: Giselbert Hinz, Abteilung für Strukturelle Zellphysiologie, Albrecht-von-Haller Institut für Pflanzenwissenschaften, Georg-August Universität, Untere Karspüle 2, 37073-Göttingen, Germany. Tel:49-551-397862

Developing pea cotyledons contain functionally different vacuoles,a protein storage vacuole and a lytic vacuole. Lumenal as wellas membrane proteins of the protein storage vacuole exit theGolgi apparatus in dense vesicles rather than in clathrin-coatedvesicles (CCVs). Although the sorting receptor for vacuolarhydrolases BP-80 is present in CCVs, it is not detectable indense vesicles. To localize these different vacuolar sortingevents in the Golgi, we have compared the distribution of vacuolarstorage proteins and of ${alpha}$ -TIP, a membrane protein of the proteinstorage vacuole, with the distribution of the vacuolar sortingreceptor BP-80 across the Golgi stack. Analysis of immunogoldlabeling from cryosections and from high pressure frozen sampleshas revealed a steep gradient in the distribution of the storageproteins within the Golgi stack. Intense labeling for storageproteins was registered for the cis-cisternae, contrasting withvery low labeling for these antigens in the trans-cisternae.The distribution of BP-80 was the reverse, showing a peak inthe trans-Golgi network with very low labeling of the cis-cisternae.These results indicate a spatial separation of different vacuolarsorting events in the Golgi apparatus of developing pea cotyledons.

Key Words: Golgi apparatus, clathrin-coated vesicle, dense vesicles, storageprotein, vacuolar sorting receptor

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