Stabilization of Synthetic Collagen Triple Helices: Charge Pairs and Covalent Capture.

Collagen mimetic peptides are composed of triple helices. Triple helical formation frequently utilizes charge pair interactions to direct protein assembly. The design of synthetic triple helices is challenging due to the large number of competing species and the overall fragile nature of collagen mimetics. A successfully designed triple helix incorporates both positive and negative criteria to achieve maximum specificity of the supramolecular assembly. Intrahelical charge pair interactions, particularly those involved in lysine-aspartate and lysine-glutamate pairs, have been especially successful both in driving helix specificity and for subsequent stabilization by covalent capture. Despite this progress, the important sequential and geometric relationships of charged residues in a triple helical context have not been fully explored for either supramolecular assembly or covalent capture stabilization. In this study, we compare the eight canonical axial and lateral charge pairs of lysine and arginine with glutamate and aspartate to their noncanonical, reversed charge pairs. These findings are put into the context of collagen triple helical design and synthesis.

Beyond the Triple Helix: Exploration of the Hierarchical Assembly Space of Collagen-like Peptides.

The de novo design of self-assembling peptides has garnered significant attention in scientific research. While alpha-helical assemblies have been extensively studied, exploration of polyproline type II (PPII) helices, such as those found in collagen, remains relatively limited. In this study, we focused on understanding the sequence-structure relationship in hierarchical assemblies of collagen-like peptides, using defense collagen SP-A as a model. By dissecting the sequence derived from SP-A and synthesizing short collagen-like peptides, we successfully constructed a discrete bundle of hollow triple helices. Mutation studies pinpointed amino acid sequences, including hydrophobic and charged residues that are critical for oligomer formation. These insights guided the de novo design of collagen-like peptides, resulting in the formation of diverse quaternary structures, including discrete and heterogenous bundled oligomers, 2D nanosheets, and pH-responsive nanoribbons. Our study represents a significant advancement in the understanding and harnessing of collagen higher-order assemblies beyond the triple helix.

Uncovering hypercoagulation status using rotational thromboelastometry in patients with sepsis presented with hypocoagulation based on conventional coagulation tests: an observational study.

OBJECTIVE: Conventional coagulation tests (CCTs) cannot identify hypercoagulation, despite being common in patients with sepsis. Moreover, CCTs overdiagnose hypocoagulation, which increases unnecessary blood transfusion. Therefore, we aimed to use rotational thromboelastometry (ROTEM) to classify the coagulation status of patients with sepsis with abnormal CCTs and to identify the main coagulation components that affect coagulation status. PATIENTS AND METHODS: This study was part of an observational study to investigate ROTEM use in 161 patients with sepsis with the Sepsis-3 criteria. They underwent concurrent CCTs and ROTEM assessments within 24 hours of Intensive Care Unit admission at the University Medical Center, Ho Chi Minh City, from June 2020 to December 2021. This study only extracted data from patients with sepsis with abnormal CCTs, including activated partial thromboplastin time ratio, international normalized ratio (INR), platelet count, and fibrinogen concentration. RESULTS: A total of 158 patients with sepsis with abnormal CCTs had a median age of 69, and 48.7% were women. Of 34 patients with INR ≥1.6, ROTEM identified 11.8% with hypercoagulation and 20.6% with normal coagulation. Of 29 patients with platelet counts <100 (103/mm3), ROTEM identified 3.5% with hypercoagulation and 24.1% with normal coagulation. In the ROTEM-based hypercoagulability group, an increase in maximum clot firmness was observed in 95.1% of cases; also, this group had significantly higher plasma fibrinogen concentrations than other groups (p<0.005). CONCLUSIONS: ROTEM can reveal hypercoagulability in patients with sepsis with hypocoagulation based on CCTs. Hyperfibrinogenemia causes hypercoagulation in patients with sepsis.