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.