Apo alpha-lactalbumin and lysozyme are colocalized in their subsequently formed spherical supramolecular assembly.
We have reported previously that the calcium-depleted form of bovine alpha-lactalbumin (apo alpha-LA) interacts with hen egg-white lysozyme (LYS) to form spherical supramolecular structures. These supramolecular structures contain an equimolar ratio of the two proteins. We further explore here the organization of these structures. The spherical morphology and size of the assembled LYS/apo alpha-LA supramolecular structures were demonstrated using confocal scanning laser microscopy and scanning electron microscopy. From confocal scanning laser microscopy experiments with labelled proteins, it was found that LYS and apo alpha-LA were perfectly colocalized and homogeneously distributed throughout the entire three-dimensional structure of the microspheres formed. The spatial colocalization of the two proteins was also confirmed by the occurrence of a fluorescence resonance energy transfer phenomenon between labelled apo alpha-LA and labelled LYS. Polarized light microscopy analysis revealed that the microspheres formed differ from spherulites, a higher order semicrystalline structure. As the molecular mechanism initiating the formation of these microspheres is still unknown, we discuss the potential involvement of a LYS/apo alpha-LA heterodimer as a starting block for such a supramolecular assembly.
Nigen
Michaël
M
INRA, Agrocampus Rennes, France.
Croguennec
Thomas
T
Madec
Marie-Noëlle
MN
Bouhallab
Saïd
S
D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D002417:Cattle; D002645:Chickens; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D007768:Lactalbumin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D008859:Microscopy, Polarization; D008863:Microspheres; D046912:Multiprotein Complexes / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D011485:Protein Binding
D001059:Apoproteins; D005456:Fluorescent Dyes; D046912:Multiprotein Complexes; C041966:apo-alpha-lactalbumin; D007768:Lactalbumin; D009113:Muramidase; D016650:Fluorescein-5-isothiocyanate