α-Lactalbumin (α-LA) is a small (Mr 14,200), acidic (pI 4-5), Ca2+-binding protein. α-LA is a regulatory component of lactose synthase enzyme system. α-LA is very important in infant nutrition since it constitutes a large part of the whey and total protein in human milk. The protein possesses a single strong Ca2+-binding site, which can also bind Mg2+, Mn2+, Na+, K+, and some other metal cations. It contains several distinct Zn2+-binding sites. Physical properties of α-LA strongly depend on the occupation of its metal binding sites by metal ions. In the absence of bound metal ions α-LA is in the molten globule-like state. The binding of metal ions, and especially of Ca2+, increases stability of α-LA against action of heat, various denaturing agents, and proteases, while the binding of Zn2+ to the Ca2+-loaded protein decreases its stability and causes its aggregation. The thermal unfolding of apo-α-LA takes place in the temperature region from 10 to 30 °C. The binding of Ca2+ under the conditions of low ionic strength shifts the thermal transition to higher temperatures by more than 40 °C. The binding of Mg2+, Na+, and K+ increases protein stability as well. The stronger an ion binds to the protein, the more pronounced the thermal transition shift. All four classes of surfactants (anionic, cationic, non-ionic, and zwitterionic) denature α-LA and the denaturation involves at least one intermediate. The position of any denaturation transition in α-LA (half-transition temperature, half-transition pressure, half-transition denaturant concentration) depends upon metal ion concentration in solution (especially if this metal ion is Ca2+). Therefore, values of denaturation temperature or urea or guanidine hydrochloride denaturing concentration are relatively meaningless for α-LA without specifying the metal ion content(s) and their solution concentration(s). At a neutral or slightly acidic pH at a physiological temperature, α-LA can associate with membranes. The conformations of the membrane-bound protein range from native-like to molten globule-like states. At a low pH, α-LA penetrates the interior of the negatively charged membranes and exhibits a molten globule conformation. Depending on external conditions, α-LA can form amyloid fibrils, amorphous aggregates, nanoparticles, and nanotubes. At pH 2, α-LA in the classical molten globule conformation can form amyloid fibrils. Some of these aggregated states of α-LA (nanoparticles, nanotubes) can be used in practical applications such as drug delivery to tissues and organs. The structure and self-assembly behavior of α-LA are governed by a subtle balance between hydrophobic and polar interactions and this balance can be finely tuned through the addition of selected substances. Small size nanoparticles of α-LA (100 to 200 nm) can be obtained with the use of various desolvating agents. Partially hydrolyzed α-LA can form nanotubes. α-LA and some of its fragments possess bactericidal and antiviral activities. Complexes of partially unfolded α-LA with oleic acid showed significant cytotoxicity to various tumor and bacterial cells. α-LA in such complexes plays a role of a delivery carrier of cytotoxic fatty acid molecules into tumor cells across the cell membrane. Cytotoxic protein–oleic acid complexes possess a common core-shell structure, where an oily core is made of a micellar oleic acid, whereas a proteinaceous shell, which stabilizes the oleic acid micelle, is formed from the flexible, partially unfolded proteins. These complexes called liprotides (lipids and partially denatured proteins), which are potential novel anti-tumorous drugs, can be considered as molten globular containers filled with the toxic oil.

α-Lactalbumin (α-LA) is a small (Mr 14,200), acidic (pI 4-5), Ca²⁺-binding protein. α-LA is a regulatory component of lactose synthase enzyme system. α-LA is very important in infant nutrition since it constitutes a large part of the whey and total protein in human milk. The protein possesses a single strong Ca²⁺-binding site, which can also bind Mg²⁺, Mn²⁺, Na⁺, K⁺, and some other metal cations. It contains several distinct Zn²⁺-binding sites. Physical properties of α-LA strongly depend on the occupation of its metal binding sites by metal ions. In the absence of bound metal ions α-LA is in the molten globule-like state. The binding of metal ions, and especially of Ca²⁺, increases stability of α-LA against action of heat, various denaturing agents, and proteases, while the binding of Zn²⁺ to the Ca²⁺-loaded protein decreases its stability and causes its aggregation. The thermal unfolding of apo-α-LA takes place in the temperature region from 10 to 30 °C. The binding of Ca²⁺ under the conditions of low ionic strength shifts the thermal transition to higher temperatures by more than 40 °C. The binding of Mg²⁺, Na⁺, and K⁺ increases protein stability as well. The stronger an ion binds to the protein, the more pronounced the thermal transition shift. All four classes of surfactants (anionic, cationic, non-ionic, and zwitterionic) denature α-LA and the denaturation involves at least one intermediate. The position of any denaturation transition in α-LA (half-transition temperature, half-transition pressure, half-transition denaturant concentration) depends upon metal ion concentration in solution (especially if this metal ion is Ca²⁺). Therefore, values of denaturation temperature or urea or guanidine hydrochloride denaturing concentration are relatively meaningless for α-LA without specifying the metal ion content(s) and their solution concentration(s). At a neutral or slightly acidic pH at a physiological temperature, α-LA can associate with membranes. The conformations of the membrane-bound protein range from native-like to molten globule-like states. At a low pH, α-LA penetrates the interior of the negatively charged membranes and exhibits a molten globule conformation. Depending on external conditions, α-LA can form amyloid fibrils, amorphous aggregates, nanoparticles, and nanotubes. At pH 2, α-LA in the classical molten globule conformation can form amyloid fibrils. Some of these aggregated states of α-LA (nanoparticles, nanotubes) can be used in practical applications such as drug delivery to tissues and organs. The structure and self-assembly behavior of α-LA are governed by a subtle balance between hydrophobic and polar interactions and this balance can be finely tuned through the addition of selected substances. Small size nanoparticles of α-LA (100 to 200 nm) can be obtained with the use of various desolvating agents. Partially hydrolyzed α-LA can form nanotubes. α-LA and some of its fragments possess bactericidal and antiviral activities. Complexes of partially unfolded α-LA with oleic acid showed significant cytotoxicity to various tumor and bacterial cells. α-LA in such complexes plays a role of a delivery carrier of cytotoxic fatty acid molecules into tumor cells across the cell membrane. Cytotoxic protein–oleic acid complexes possess a common core-shell structure, where an oily core is made of a micellar oleic acid, whereas a proteinaceous shell, which stabilizes the oleic acid micelle, is formed from the flexible, partially unfolded proteins. These complexes called liprotides (lipids and partially denatured proteins), which are potential novel anti-tumorous drugs, can be considered as molten globular containers filled with the toxic oil.

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