Abstract: This disclosure describes a method for de novo bottom-up assembly and rational design of allosteric biosensors with programmable input-output behaviors that respond to soluble factors selectively enriched in tumors and trigger co-stimulation and cytokine signals. The disclosed method of effective mechanical coupling and biosensor signaling potency correlates with anti-tumor function. This disclosure provides synthetic biosensors with custom-built sensing and responses for basic and translational cell engineering applications.

Abstract: The present invention relates to a computer-implemented method for engineering the interaction between a protein and a cognate peptide that are capable of forming a molecular complex, wherein the method comprises (a) preparing in silico a library of test peptides based on the cognate peptide, and the molecular complex of the protein and the cognate peptide; and/or (a?) preparing in silico a library of test protein scaffolds based at least on the parts of the protein that participate in forming the molecular complex with the cognate peptide; (b) docking in silico (i) the library of test peptides onto the library of protein scaffolds by modelling peptide-protein molecular complexes, or (ii) the cognate peptide on the library of protein scaffolds, or the library of test peptides onto the protein or the parts of the protein that participate in forming the molecular complex with the cognate peptide by modelling peptide-protein molecular complexes; (c) identifying the test peptides in the library and/or the protein

Abstract: The present disclosure relates to a method for generating variants of a protein based on a native protein regulated by allosteric pathway, the method comprising: i) providing 3D structures of the native protein; ii) identifying at least one pair of coupled allosteric sites within the amino acid sequence of the native protein named microswitch; iii) generating in silico mutations of said identified microswitch to generate a pool of variants; iv) computing at least one score reflecting the variation in allosteric coupling; and/or the variation in the relative stability v) predicting the activity of each variant compared to the native protein based on the computed score. The disclosure also concern a computer implemented program to carry out said method, and a variant of a protein or an active fragment thereof, a polynucleotide.

Abstract: A transmission is provided with a park pawl moveable around an axis and positioned over a park gear. An actuator assembly is provided for pushing the park pawl against the effect of a return spring in a blocked position thus blocking the park gear. In order to provide a transmission in which the risk of an erroneous engagement of the park pawl due to a broken spring is avoided, means, such as a mechanical coupling, are provided for mechanically coupling the park pawl to the actuator when the park pawl is in unblocked position. Due to this mechanical coupling, the park pawl is retained in the unblocked position and prevented from falling into the park gear even if the return spring breaks.