We used atomic force microscopy (AFM) to characterize the plasma membrane of Xenopus laevis oocytes. The samples were prepared according to novel protocols, which allowed the investigation of the extra- and intracellular sides of the membrane, both of which showed sparsely distributed spherical-like protrusions. Regions with comparably sized and densely packed structures arranged in an orderly manner were visualized and dimensionally characterized. In particular, two different arrangements, hexagonal and square packing, were recognizable in ordered regions. The lateral dimension of structures visualized on the external side had a normal distribution centered on 25.5 (plus or minus) 0.3 nm (mean value (plus or minus) SE), whereas that on the intracellular side showed a normal distribution centered on 30.2 (plus or minus) 0.8 nm. The height of the protrusions was 2-5 nm on the external side and 1-3 nm on the intracellular side. The mean number of structures on the external and intracellular sides of the plasma membrane was about 1000 (mu)(-2) and 850 (mu)(-2) respectively. Trypsin treatment greatly decreased the size of the membrane protrusions, thus confirming the proteic nature of the structures. These results show that AFM is a useful tool for structural characterization of proteins in a native eukaryotic membrane

Atomic force microscopy characterization of Xenopus laevis oocyte plasma membrane / F. Orsini, M. Santacroce, C. Perego, C. Lenardi, M. Castagna, S.A. Mari, V.F. Sacchi, G. Poletti. - In: MICROSCOPY RESEARCH AND TECHNIQUE. - ISSN 1059-910X. - 69:10(2006), pp. 826-834. [10.1002/jemt.20353]

Atomic force microscopy characterization of Xenopus laevis oocyte plasma membrane

F. Orsini
Primo
;
M. Santacroce
Secondo
;
C. Perego;C. Lenardi;M. Castagna;S.A. Mari;V.F. Sacchi
Penultimo
;
G. Poletti
Ultimo
2006

Abstract

We used atomic force microscopy (AFM) to characterize the plasma membrane of Xenopus laevis oocytes. The samples were prepared according to novel protocols, which allowed the investigation of the extra- and intracellular sides of the membrane, both of which showed sparsely distributed spherical-like protrusions. Regions with comparably sized and densely packed structures arranged in an orderly manner were visualized and dimensionally characterized. In particular, two different arrangements, hexagonal and square packing, were recognizable in ordered regions. The lateral dimension of structures visualized on the external side had a normal distribution centered on 25.5 (plus or minus) 0.3 nm (mean value (plus or minus) SE), whereas that on the intracellular side showed a normal distribution centered on 30.2 (plus or minus) 0.8 nm. The height of the protrusions was 2-5 nm on the external side and 1-3 nm on the intracellular side. The mean number of structures on the external and intracellular sides of the plasma membrane was about 1000 (mu)(-2) and 850 (mu)(-2) respectively. Trypsin treatment greatly decreased the size of the membrane protrusions, thus confirming the proteic nature of the structures. These results show that AFM is a useful tool for structural characterization of proteins in a native eukaryotic membrane
AFM; Membrane proteins; Plasma membrane; Xenopus laevis oocytes
Settore BIO/09 - Fisiologia
Settore FIS/01 - Fisica Sperimentale
2006
http://www.ncbi.nlm.nih.gov/entrez/utils/fref.fcgi?itool=AbstractPlus-def&PrId=3058&uid=16886228&db=pubmed&url=http://dx.doi.org/10.1002/jemt.20353
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/23217
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