In Paramecium primaurelia mating interactions take place immediately after mixing mating-competent cells of opposite mating types. The cells clump in clusters (mating reaction) and then separate in pairs. Previous results have shown that sialic acid-containing glycoconjugates are present on the cell surface and are involved in mating-cell pairing. In order to identify the sialic acid-containing glycoprotein(s), we first metabolically radiolabelled non-mating competent cells with D-[6-3H]galactose, and then analyzed the radiolabelled proteins by anion exchange chromatography. We characterized a 38 kDa (gp38) sialic acid-containing glycoprotein and raised the corresponding polyclonal antibody by means of which we localized the antigen at the level of the oral region of non-mating competent cells and on the ciliary surface of mating-competent cells. Immunoblot analysis of the ciliary protein fraction showed that the anti-gp38 serum interacted with a 38 kDa protein in both mating types I and II cells. We also demonstrated the functional activity of gp38 in the mating reaction by means of anti-gp38 antibody competition assays.

M., O., DELLA GIOVAMPAOLA, C., F., T., Focarelli, R., Rosati, F., M. U., D.C. (2008). Identification and characterization of a 38 kDa glycoprotein functionally associated with mating activity of Paramecium primaurelia. EUROPEAN JOURNAL OF PROTISTOLOGY, 44, 81-90 [10.1016/j.ejop.2007.07.002].

Identification and characterization of a 38 kDa glycoprotein functionally associated with mating activity of Paramecium primaurelia

DELLA GIOVAMPAOLA, CINZIA;FOCARELLI, RICCARDO;ROSATI, FLORIANA;
2008-01-01

Abstract

In Paramecium primaurelia mating interactions take place immediately after mixing mating-competent cells of opposite mating types. The cells clump in clusters (mating reaction) and then separate in pairs. Previous results have shown that sialic acid-containing glycoconjugates are present on the cell surface and are involved in mating-cell pairing. In order to identify the sialic acid-containing glycoprotein(s), we first metabolically radiolabelled non-mating competent cells with D-[6-3H]galactose, and then analyzed the radiolabelled proteins by anion exchange chromatography. We characterized a 38 kDa (gp38) sialic acid-containing glycoprotein and raised the corresponding polyclonal antibody by means of which we localized the antigen at the level of the oral region of non-mating competent cells and on the ciliary surface of mating-competent cells. Immunoblot analysis of the ciliary protein fraction showed that the anti-gp38 serum interacted with a 38 kDa protein in both mating types I and II cells. We also demonstrated the functional activity of gp38 in the mating reaction by means of anti-gp38 antibody competition assays.
M., O., DELLA GIOVAMPAOLA, C., F., T., Focarelli, R., Rosati, F., M. U., D.C. (2008). Identification and characterization of a 38 kDa glycoprotein functionally associated with mating activity of Paramecium primaurelia. EUROPEAN JOURNAL OF PROTISTOLOGY, 44, 81-90 [10.1016/j.ejop.2007.07.002].
File in questo prodotto:
File Dimensione Formato  
http pdn.sciencedirect.pdf

non disponibili

Tipologia: Post-print
Licenza: NON PUBBLICO - Accesso privato/ristretto
Dimensione 513.25 kB
Formato Adobe PDF
513.25 kB Adobe PDF   Visualizza/Apri   Richiedi una copia

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11365/21316
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo