RESUMO
We investigated the growth of biosynthetic protein polymers with templated curvature on pluronic nanospheres. The protein has a central silk-like block containing glutamic residues (S(E)) and collagen-like end-blocks (C). The S(E) blocks stack into filaments when their charge is removed (pH <5). Indeed, at low pH curved and circular fibers are formed at the surface of the nanospheres, which keep their shape after removal of the pluronics. The data reveal the mechanism of the templated fibril-growth: The growth of protein assemblies is nucleated in solution; small protein fibrils adsorb on the nanospheres, presumably due to hydrogen bond formation between the silk-like blocks and the pluronic PEO blocks. The surface of the pluronic particles templates further growth. At relatively low protein/pluronic weight ratios, only a fraction of the nanospheres bears protein fibers, pointing to a limiting amount of nuclei in solution. Because the nanospheres capture fibrils at an early stage of growth, they can be used to separate growth and nucleation rates in protein fibril formation. Moreover, the nanoparticle-templated growth of stable curved fibers opens ways to build proteinaceous nanocapsules from designed protein polymers.
Assuntos
Colágeno/química , Nanosferas/química , Poloxâmero/química , Polímeros/química , Seda/química , Matriz Extracelular , Humanos , Propriedades de SuperfícieRESUMO
An asymmetric ('hybrid') triblock polypeptide TR4H with two different, orthogonally self-assembling end blocks has been constructed by conjugating a long (37 kDa) random coil block (R4) with a triple helix former T = (Pro-Gly-Pro)9 at the N terminus, and a histidine hexamer ('Histag', H) at the C terminus. This molecule can form trimers at room temperature by assembly of the T blocks, which can in turn assemble upon addition of Ni(2+), by association of Ni complexes involving the H block. This results in reversible hydrogels with dual responsiveness. We have studied mechanical properties of these gels, and compared them to gels formed by the symmetric triblock TR8T which is equivalent to a dimer of TR4H, but can only form triple helix-based networks. We find that there is an optimum mole ratio for Ni(2+) with respect to the polypeptide of about 1; gels are weaker at both lower and higher Ni(2+) dose. At the optimum dose, the high-frequency storage modulus is in between the value expected for nickel-induced dimerization and trimerization of the H blocks. We also find that the gels relax on time scales of about 50 s, which is two orders of magnitude faster than for TR8T gels, implying that relaxation is dominated by the dynamics of the Ni(2+) complex.
RESUMO
Recombinant protein polymers, which can combine different bioinspired self-assembly motifs in a well-defined block sequence, have large potential as building blocks for making complex, hierarchically structured materials. In this paper we demonstrate the stepwise formation of thermosensitive hydrogels by combination of two distinct, orthogonal self-assembly mechanisms. In the first step, fibers are coassembled from two recombinant protein polymers: (a) a symmetric silk-like block copolymer consisting of a central silk-like block flanked by two soluble random coil blocks and (b) an asymmetric silk-collagen-like block copolymer consisting of a central random-coil block flanked on one side by a silk-like block and on the other side a collagen-like block. In the second step, induced by cooling, the collagen-like blocks form triple helices and thereby cross-link the fibers, leading to hydrogels with a thermo-reversibly switchable stiffness. Our work demonstrates how complex self-assembled materials can be formed through careful control of the self-assembly pathway.
Assuntos
Colágeno/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Proteínas Recombinantes/síntese química , Seda/química , Sequência de Aminoácidos , Colágeno/síntese química , Hidrogel de Polietilenoglicol-Dimetacrilato/síntese química , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Polímeros/síntese química , Polímeros/química , Proteínas Recombinantes/química , Seda/síntese química , TemperaturaRESUMO
In this report, we study the self-assembly of two silk-elastin-like proteins: one is a diblock S(24)E(40) composed of 24 silk-like (S) repeats and 40 elastin-like (E) repeats; the other is a triblock S(12)C(4)E(40), in which the S and E blocks are separated by a random coil block (C(4)). Upon lowering the pH, the acidic silk-like blocks fold and self-assemble into fibrils by a nucleation-and-growth process. While silk-like polymers without elastin-like blocks form fibrils by heterogeneous nucleation, leading to monodisperse populations, the elastin-like blocks allow for homogeneous nucleation, which gives rise to polydisperse length distributions, as well as a concentration-dependent fibril length. Moreover, the elastin-like blocks introduce temperature sensitivity: at high temperature, the fibrils become sticky and tend to bundle and aggregate in an irreversible manner. Concentrated solutions of S(12)C(4)E(40) form weak gels at low pH that irreversibly lose elasticity in temperature cycling; this is also attributed to fibril aggregation.