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When the K + channel-like protein Kesv from Ectocarpus siliculosus virus 1 is heterologously expressed in mammalian cells, it is sorted to the mitochondria. This targeting can be redirected to the endoplasmic reticulum (ER) by altering the codon usage in distinct regions of the gene or by inserting a triplet of hydrophobic amino acids (AAs) into the protein's C-terminal transmembrane domain (ct-TMD). Systematic variations in the flavor of the inserted AAs and/or its codon usage show that a positive charge in the inserted AA triplet alone serves as strong signal for mitochondria sorting. In cases of neutral AA triplets, mitochondria sorting are favored by a combination of hydrophilic AAs and rarely used codons; sorting to the ER exhibits the inverse dependency. This propensity for ER sorting is particularly high when a common codon follows a rarer one in the AA triplet; mitochondria sorting in contrast is supported by codon uniformity. Since parameters like positive charge, hydrophobic AAs, and common codons are known to facilitate elongation of nascent proteins in the ribosome the data suggest a mechanism in which local changes in elongation velocity and co-translational folding in the ct-TMD influence intracellular protein sorting., (© 2023 The Authors. Traffic published by John Wiley & Sons Ltd.)

To elucidate the effects of extracellular polymeric substance (EPS) on the sludge dewaterability, this study comparatively investigated the changes in EPS composition and spatial distribution, together with the sludge dewaterability after lysozyme (LZM) conditioning. The protein concentration in the tightly bound EPS (TB-EPS) increased from 3.47 mg g -1 DS to 4.99 mg g -1 DS within the first 2 min, then gradually decreased, which could be described by a piecewise linear function. Unlike TB-EPS, the protein content variation trend in both soluble EPS (S-EPS) and loosely bound EPS (LB-EPS) followed the typical first-order kinetics. Additionally, the extended DLVO theory was employed in combination with viscoelastic acoustic response analysis to further explore the impact of EPS composition on water adhesion and microbial cell. After the extraction of S-EPS from the conditioned sludge, the adsorption free energy (ΔG adh ) of EPS ascended to -61.05 mJ m -2 , indicating the weakened microbial hydrophobicity. By contrast, the ΔG adh value declined after the subsequent extraction of LB-EPS and TB-EPS. Meanwhile, the adsorption potential energy between S-EPS and microbial cells showed an increasing trend, whereas the repulsion potential energy between TB-EPS and microbial cells fell to 1.40 × 10 4  kT, signifying a weakened adsorption capacity to water. Accordingly, the viscosity and shear modulus of each EPS layer were reduced after conditioning, which contributed to the transformation of bound water into free water. These changes reasonably explained the results that the water content in the dewatered sludge after conditioning was reduced to 58.54%, and the bound water content decreased by 15.06%., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)