Backbone Assignment of the MALT1 Paracaspase by Solution NMR.

Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) is a unique paracaspase protein whose protease activity mediates oncogenic NF-κB signalling in activated B cell-like diffuse large B cell lymphomas (ABC-DLBCLs). ABC-DLBCLs are aggressive lymphomas with high resistance to current chemotherapies. Low survival rate among patients emphasizes the urgent need for alternative treatment options. The characterization of the MALT1 will be an essential tool for developing new target-directed drugs against MALT1 dependent disorders. As the first step in the atomic-level NMR studies of the system, here we report, the (15)N/(13)C/(1)H backbone assignment of the apo form of the MALT1 paracaspase region together with the third immunoglobulin-like (Ig3) domain, 44 kDa, by high resolution NMR. In addition, the non-uniform sampling (NUS) based targeted acquisition procedure is evaluated as a mean of decreasing acquisition and analysis time for larger proteins

Mapping of glutathione transferase (GST) genes in the rat.

Glutathione transferases (GST) make up a large group of related enzymes in mammalian tissues. The enzyme molecules are dimeric and at least 13 different subunits occur in the rat. Each subunit appears to be coded for by a distinct gene, and thus there is a large GST gene family in the rat. Recently, there have been several reports of the mapping of rat GST genes. In the present communication we confirm the previous assignments and extend the data with the mapping to rat chromosome 2 of a previously unmapped GST gene (Gstm1), and with the regional mapping of seven Gstp genes. These mappings provide further evidence for conservation of syntenic gene relationships among mammals. The human homologs of Gstm1 map to chromosome 1, and belong to a group of 9 genes that show conserved synteny on rat chromosome 2. The corresponding murine genes in most cases map to mouse chromosome 3. Similarly, the human homolog of Gstp maps to chromosome 11, and is one of 10 genes that exhibit conserved synteny on rat chromosome 1. The corresponding mouse genes map to mouse chromosome 7. Previously only one gene on rat chromosome 8 had a human homolog on chromosome 6, and rat Gsta1 is the second instance. Based on these mappings it appears that a new group of genes will exhibit conserved synteny on rat chromosome 8, human chromosome 6 and mouse chromosome 9. Interestingly, each of the three groups of conserved synteny seems to span the region across the centromeres of the human chromosomes.Comparative StudyJournal ArticleResearch Support, Non-U.S. Gov'tSCOPUS: ar.jinfo:eu-repo/semantics/publishe

Estimate of the secondary structure in MALT1_Casp-Ig3(338–719).

<p>Secondary chemical shifts (Δδ) were calculated by subtracting random coil chemical shifts corrected for nearest-neighbour effects from ¹³C’, ¹³Cα and ¹³Cβ chemical shifts corrected for deuterium isotope shifts. Consecutive values above 0.7 indicates alpha helix, while consecutive values below -0.7 indicates beta strand for Δδ¹³C’ and Δδ¹³Cα. The opposite is true for Δδ¹³Cβ. The CSI for the three nuclei were averaged and reported as a “consensus” CSI. β3, β3A and β3B are denoted β3 AB in the Fig. The star (*) indicates that the secondary structure is part of the Ig3 domain.</p

¹H-¹⁵N TROSY spectrum of MALT1_Casp-Ig3(338–719) with the assigned amino acid residue number annotated.

<p>¹H-¹⁵N TROSY spectrum of MALT1_Casp-Ig3(338–719) with the assigned amino acid residue number annotated.</p

Peak appearance progress during the course of the TA procedure for the MALT1 sample.

<p>The horizontal axis shows the total measurement time excluding the HNCO experiment, which was recorded prior to the TA. The spectral processing and analysis was done automatically during the course of the data acquisition.</p

Estimated secondary structure from NMR experiments (in black) compared to secondary structure from the in-house X-ray structure (in dark grey) and from the published X-ray structure of <i>apo</i> MALT1_Casp-Ig3, PDB ID: 3V55 (in light grey).

<p>Alpha helices are indicated with a greater symbol size than the beta-sheets. β3, β3A and β3B are denoted β3 AB in the Fig. The star (*) indicates that the secondary structure is part of the Ig3 domain.</p