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Two of the peripheral stalks protrude from subunit C, one from the "head" (residues 166 263) and "foot" domains (residues 1 48 and 324 373), while the third is seen projecting from the amino-terminus of subunit H.
Two of these formations are occupied by the "head" (residues 166 263) and a "foot" (residues 1 48 and 324 373) [30] domains of subunit C, while the third is populated by the amino-termini of subunit H which also serves as the attachment point for one of the peripheral stalks.
Talin (2541 amino acids) is composed of a globular head (residues 1 400), containing a FERM domain, connected to a flexible rod (residues 482 2541) by a short linker sequence containing a calpain-II cleavage site (Critchley, 2004).
However, expression of the whole talin head (residues 1 405) was able to support β1A-integrin activation, and this required residues 1 85, which precede the FERM domain (referred to as the F0 domain), plus the F1 FERM domain and the integrin-binding F3 domain (Bouaouina et al, 2008).
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It consists of 1066 amino acids, has a molecular weight of 117 kDa and, based on its protein structure, can be divided into a head region (residues 1 835) and a tail region (residues 896 1066) connected by a proline-rich region and a flexible hinge (residues 836 895) (10).
We additionally constructed and purified a monomeric Kar3 head encompassing residues 353 729 with an N-terminal Halo-Tag.
As suggested, we have tested the motility of a monomeric Kar3 head construct (residues 353-729 fused to an N-terminal Halo tag).
(I ) Typical kymographs of a monomeric Kar3 head construct (residues 353 729 fused to an N-terminal Halo tag) in the presence of ATP.
Using the assignments of F1, it was possible to identify the resonances of the F1 loop in the [H,N]-HSQC spectrum of the whole talin head (F0F1F2F3, residues 1 400) (Supplementary Figure 4).
Previous work regarding the phosphorylation of K19 demonstrated that head domain residue serine-35 is a major phosphorylation site [20] and that K19 expressed by various cell lines and primary mouse colon epithelial cells undergoes tyrosine phosphorylation upon treatment with pervanadate, a potent tyrosine phosphatase inhibitor [21].
The large quantities of by-products generated have great potential for becoming value-added compounds, such as lipids, proteins, amino acids and enzymes, which can be extracted from wastewaters and from solid residues (head, viscera, skin, tails and flesh) that are leftovers during the canning process.
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