Recombinant production of self-assembling ß-structured peptides using SUMO as a fusion partner.

BACKGROUND: Self-assembling peptides that form nanostructured hydrogels are important biomaterials for tissue engineering scaffolds. The P11-family of peptides includes, P11-4 (QQRFEWEFEQQ) and the complementary peptides P11-13 (EQEFEWEFEQE) and P11-14 (QQOrnFOrnWOrnFOrnQQ). These form self-supporting hydrogels under physiological conditions (pH 7.4, 140 mM NaCl) either alone (P11-4) or when mixed (P11-13 and P11-14). We report a SUMO-peptide expression strategy suitable for allowing release of native sequence peptide by SUMO protease cleavage. RESULTS: We have expressed SUMO-peptide fusion proteins from pET vectors by using autoinduction methods. Immobilised metal affinity chromatography was used to purify the fusion protein, followed by SUMO protease cleavage in water to release the peptides, which were recovered by reverse phase HPLC. The peptide samples were analysed by electrospray mass spectrometry and self-assembly was followed by circular dichroism and transmission electron microscopy. CONCLUSIONS: The fusion proteins were produced in high yields and the ß-structured peptides were efficiently released by SUMO protease resulting in peptides with no additional amino acid residues and with recoveries of 46% to 99%. The peptides behaved essentially the same as chemically synthesised and previously characterised recombinant peptides in self-assembly and biophysical assays.

Effects of Two Competitive Soccer Matches on Landing Biomechanics in Female Division I Soccer Players.

Fatigue has been proposed to increase the risk of knee injury. This study tracked countermovement jump, knee isometric strength, and kinetics and kinematics in 8 female soccer players (experimental group) during an anticipated sidestep maneuver before and after two matches played over a 43-h period. Time points were: Before and after match 1 (T0 and T1), 12 h after the first match (T2), and immediately after the second match (T3). A control group participated only in practice sessions. Isometric knee extension strength decreased by 14.8% at T2 (p = 0.003), but knee flexion was not affected until T3, declining by 12.6% (p = 0.018). During the sidestep maneuver, knee joint degrees of flexion at initial contact was increased by 17.1% at T3, but maximum knee and hip angle at initial contact were unchanged. Peak resultant ground reaction force (GRF) increased by 12.6% (p = 0.047) at T3 (3.03 xBW) from 2.69 xBW at T0, while posterior GRF was significantly higher than T0 at all three subsequent time points (T1 = 0.82 ± 0.23 xBW, T2 = 0.87 ± 0.22 xBW, T3 = 0.89 ± 0.22 xBW). Anterior tibial shear force increased significantly (p = 0.020) at T3 (1.24 ± 0.12 xBW) compared to T1 (1.15 ± 0.13 xBW), an 8.8% increase. Lateral tibial shear force was significantly higher at both T1 (0.95 ± 0.20 xBW) and T3 (1.15 ± 0.38 xBW) compared to T0 (0.67 ± 0.25 xBW). These findings suggest that participation in a soccer match has significant effects on both physical performance parameters and kinetics/kinematics during a sidestep cut, but these can be more pronounced after a second match with short rest.