SiiE from Salmonella enterica is a giant 5,559-residue-long nonfimbrial adhesin that is secreted by a type 1 secretion system (T1SS) and initiates bacterial adhesion to polarized host cells. Structural insight has been gained into the 53 bacterial Ig-like (BIg) domains of SiiE, which account for 94% of the entire SiiE sequence. The crystal structure of a fragment comprising BIg domains 50 to 52 of SiiE reveals the BIg domain architecture and highlights two types of SiiE-specific Ca²⁺-binding sites. Sequence homology considerations suggest that full-length SiiE interacts with more than 100 Ca²⁺ ions. Molecular dynamics simulations and single-molecule imaging indicate that Ca²⁺ binding confers SiiE with a rigid 200 nm rod-like habitus that is required to reach out beyond the Salmonella lipopolysaccharide layer and to promote adhesion to host cells. The crystal structure suggests plausible routes for the establishment of the initial contact between Salmonella and host cells.
Salmonella infections can be life-threatening. SiiE is a giant adhesion molecule of 5559 amino acids that is encoded in Salmonella pathogenicity island 4 (SPI4) and that promotes the initial contact between the pathogen and polarized epithelial cells in the intestine of the host. Starting from an engineered deletion version of SiiE (mini-SiiE; 97 kDa), limited proteolysis was used to reproducibly generate a 30 kDa fragment that readily crystallized. Mass spectrometry hints that this fragment spans the predicted Ig domains 50-52 of SiiE. Crystals of both native and selenomethionine-labelled protein could be obtained in space group C2 and diffraction data were recorded to a resolution of 1.85 Å .
The dynamic elastic modulus (Ed) and the coefficient of wall viscosity (eta w) of the tail artery of normotensive rats were determined as functions of the circumferential wall stress under quasistatic and dynamic conditions. The experiments were performed under strong smooth muscle activation induced by norepinephrine, and during relaxation induced by papaverine. The following results were obtained. 1. Ed and eta w increase with increasing wall stress. At a given wall stress, Ed is virtually independent of frequency while eta w decreases markedly with increasing frequency. This behaviour of eta w is called thixotropy or pseudoplasticity. 2. In the wall stress range from 5--60 kPa the values of Ed, and in the wall stress range from 60--140 kPa those of eta w obtained under smooth muscle activation and during relaxation are virtually identical. 3. In the relaxed smooth muscle, the phase angles between sinusoidal pressure and radius changes area virtually independent of the mean wall stress at all frequencies. In the low stress range, the phase angles are greater at low frequencies in the activated state than in the relaxed state, decrease with increasing wall stress, and are virtually identical to the values under papaverine at high wall stresses. At high frequencies no dependence of the phase angles on the mean wall stress can be seen.
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