Structure of the Monkeypox virus profilin-like protein A42R reveals potential functional differences from cellular profilins
The infectious disease human monkeypox is spreading rapidly in 2022, causing a global health crisis. The genomics of Monkeypox virus (MPXV) have been extensively analyzed and reported, although little is known about the virus-encoded proteome. In particular, there are no reported experimental MPXV protein structures other than computational models. Here, a 1.52 Å resolution X-ray structure of the MPXV protein A42R, the first MPXV-encoded protein with a known structure, is reported. A42R shows structural similarity to profilins, which are cellular proteins that are known to function in the regulation of actin cytosk...
Source: Acta Crystallographica Section F - September 26, 2022 Category: Biochemistry Authors: Minasov, G. Inniss, N.L. Shuvalova, L. Anderson, W.F. Satchell, K.J.F. Tags: monkeypox poxviruses profilin profilin-like protein A42R actin X-ray structure emerging infectious diseases Center for Structural Genomics of Infectious Diseases research communications Source Type: research
The structure of succinyl-CoA synthetase bound to the succinyl-phosphate intermediate clarifies the catalytic mechanism of ATP-citrate lyase
Succinyl-CoA synthetase (SCS) catalyzes a three-step reaction in the citric acid cycle with succinyl-phosphate proposed as a catalytic intermediate. However, there are no structural data to show the binding of succinyl-phosphate to SCS. Recently, the catalytic mechanism underlying acetyl-CoA production by ATP-citrate lyase (ACLY) has been debated. The enzyme belongs to the family of acyl-CoA synthetases (nucleoside diphosphate-forming) for which SCS is the prototype. It was postulated that the amino-terminal portion catalyzes the full reaction and the carboxy-terminal portion plays only an allosteric role. This interpretat...
Source: Acta Crystallographica Section F - September 26, 2022 Category: Biochemistry Authors: Huang, J. Fraser, M.E. Tags: succinyl-CoA synthetase succinyl-phosphate cryogenic electron microscopy catalytic intermediates ATP-citrate lyase research communications Source Type: research
Purification, crystallization and crystallographic analysis of the PorX response regulator associated with the type IX secretion system
Pathogenic bacteria utilize specialized macromolecular secretion systems to transport virulence factors across membrane(s) and manipulate their infected host. To date, 11 secretion systems have been identified, including the type IX secretion system (T9SS) associated with human, avian and farmed-fish diseases. As a bacterial secretion system, the T9SS also facilitates gliding motility and the degradation of different macromolecules by the secretion of metabolic enzymes in nonpathogenic bacteria. PorX is a highly conserved protein that regulates the transcription of essential T9SS components and additionally mediates the fu...
Source: Acta Crystallographica Section F - September 26, 2022 Category: Biochemistry Authors: Saran, A. Weerasinghe, N. Thibodeaux, C.J. Zeytuni, N. Tags: two-component systems response regulators bacterial secretion systems bacterial pathogenesis PorX response regulator type IX secretion system research communications Source Type: research
Crystal structure of the Rho-associated coiled-coil kinase 2 inhibitor belumosudil bound to CK2 α
The small molecule belumosudil was initially identified as a selective inhibitor of Rho-associated coiled-coil kinase 2 (ROCK2) and has recently been approved for the treatment of graft-versus-host disease. However, recent studies have shown that many of the phenotypes displayed upon treatment with belumosudil were due to CK2 α inhibition. CK2 α is in itself a very promising therapeutic target for a range of conditions and has recently been put forward as a potential treatment for COVID-19. Belumosudil presents a promising starting point for the development of future CK2 α inhibitors as it provides a safe, potent and or...
Source: Acta Crystallographica Section F - September 26, 2022 Category: Biochemistry Authors: Brear, P. Hyv ö nen, M. Tags: belumosudil CK2 α inhibition Rho-associated coiled-coil kinase 2 ROCK2 inhibition kinase inhibitors selective inhibitors research communications Source Type: research
Structure of the hypothetical protein TTHA1873 from Thermus thermophilus
The crystal structure of an uncharacterized hypothetical protein, TTHA1873 from Thermus thermophilus, has been determined by X-ray crystallography to a resolution of 1.78 Å using the single-wavelength anomalous dispersion method. The protein crystallized as a dimer in two space groups: P43212 and P6122. Structural analysis of the hypothetical protein revealed that the overall fold of TTHA1873 has a β -sandwich jelly-roll topology with nine β -strands. TTHA1873 is a dimeric metal-binding protein that binds to two Ca2+ ions per chain, with one on the surface and the other stabilizing the dimeric interface of the two c...
Source: Acta Crystallographica Section F - August 30, 2022 Category: Biochemistry Authors: Yuvaraj, I. Chaudhary, S.K. Jeyakanthan, J. Sekar, K. Tags: SAD phasing hypothetical proteins metalloproteins jelly-roll topology Thermus thermophilus TTHA1873 calcium-binding proteins research communications Source Type: research
Structure of angiogenin dimer bound to double-stranded RNA
Angiogenin is an unusual member of the RNase A family and is of great interest in multiple pathological contexts. Although it has been assigned various regulatory roles, its core catalytic function is that of an RNA endonuclease. However, its catalytic efficiency is comparatively low and this has been linked to a unique C-terminal helix which partially blocks its RNA-binding site. Assuming that binding to its RNA substrate could trigger a conformational rearrangement, much speculation has arisen on the topic of the interaction of angiogenin with RNA. To date, no structural data on angiogenin – RNA interactions have been ...
Source: Acta Crystallographica Section F - August 30, 2022 Category: Biochemistry Authors: Sievers, K. Ficner, R. Tags: angiogenin ribonucleases RNP RNase A double-stranded RNA research communications Source Type: research
Crystallization and crystallographic studies of a novel chickpea 11S globulin
Chickpea is a crop that is known as a source of high-quality proteins. CL-AI, which belongs to the 11S globulin and cupin superfamily, was initially identified in chickpea seeds. CL-AI has recently been shown to inhibit various types of α -amylases. To determine its molecular mechanism, the crystal structure of CL-AI was solved at a final resolution of 2.2 Å . Structural analysis indicated that each asymmetric unit contains three molecules with threefold symmetry and a head-to-tail association, and each molecule is divided into an α -chain and a β -chain. CL-AI has high structural similarity to other 11S globulins ...
Source: Acta Crystallographica Section F - August 22, 2022 Category: Biochemistry Authors: Sun, L. Zhou, A. Zhang, F. Tags: 11S globulins chickpeas crystal structure cupin proteins CL-AI research communications Source Type: research
Crystallization and preliminary X-ray analysis of the C-type lectin domain of the spicule matrix protein SM50 from Strongylocentrotus purpuratus. Corrigendum
The identity of the crystallized protein in the article by Juneja et al. [(2014), Acta Cryst. F70, 260 – 262] is corrected. (Source: Acta Crystallographica Section F)
Source: Acta Crystallographica Section F - August 9, 2022 Category: Biochemistry Authors: Ebenhoch, R. Juneja, P. Rao, A. C ö lfen, H. Diederichs, K. Welte, W. Tags: corrigendum C-type lectin domain spicule matrix protein SM50 Strongylocentrotus purpuratus chloramphenicol acetyltransferase addenda and errata Source Type: research
Delineating the activity of the potent nicotinic acetylcholine receptor agonists (+)-anatoxin-a and ( − )-hosieine-A
The affinity and thermodynamic parameters for the interactions of two naturally occurring neurotoxins, (+)-anatoxin-a and ( − )-hosieine-A, with acetylcholine-binding protein were investigated using a fluorescence-quenching assay and isothermal titration calorimetry. The crystal structures of their complexes with acetylcholine-binding protein from Aplysia californica (AcAChBP) were determined and reveal details of molecular recognition in the orthosteric binding site. Comparisons treating AcAChBP as a surrogate for human α 4 β 2 and α 7 nicotinic acetylcholine receptors (nAChRs) suggest that the molecular features inv...
Source: Acta Crystallographica Section F - August 9, 2022 Category: Biochemistry Authors: Parker, H.P. Dawson, A. Jones, M.J. Yan, R. Ouyang, J. Hong, R. Hunter, W.N. Tags: acetylcholine-binding proteins ( )-anatoxin-a − )-hosieine-A crystal structure ligand-gated ion channels neurotoxins nicotine nicotinic acetylcholine receptors varenicline research communications Source Type: research
Crystal structures of glutamyl-tRNA synthetase from Elizabethkingia anopheles and E. meningosepticum
Elizabethkingia bacteria are globally emerging pathogens that cause opportunistic and nosocomial infections, with up to 40% mortality among the immunocompromised. Elizabethkingia species are in the pipeline of organisms for high-throughput structural analysis at the Seattle Structural Genomics Center for Infectious Disease (SSGCID). These efforts include the structure – function analysis of potential therapeutic targets. Glutamyl-tRNA synthetase (GluRS) is essential for tRNA aminoacylation and is under investigation as a bacterial drug target. The SSGCID produced, crystallized and determined high-resolution structures of...
Source: Acta Crystallographica Section F - July 28, 2022 Category: Biochemistry Authors: Brooks, L. Subramanian, S. Dranow, D.M. Mayclin, S.J. Myler, P.J. Asojo, O.A. Tags: glutamyl-tRNA synthetases undergraduate education and training Seattle Structural Genomics Center for Infectious Disease infectious diseases Elizabethkingia meningosepticum Elizabethkingia anopheles emerging infectious diseases research comm Source Type: research
Variations on a theme: crystal forms of the amino-acid transporter MhsT
The bacterial amino-acid transporter MhsT from the SLC6A family has been crystallized in complex with different substrates in order to understand the determinants of the substrate specificity of the transporter. Surprisingly, crystals of the different MhsT – substrate complexes showed interrelated but different crystal-packing arrangements. Space-group assignment and structure determination of these different crystal forms present challenging combinations of pseudosymmetry, twinning and translational noncrystallographic symmetry. (Source: Acta Crystallographica Section F)
Source: Acta Crystallographica Section F - July 26, 2022 Category: Biochemistry Authors: Neumann, C. Focht, D. Trampari, S. Lyons, J.A. Nissen, P. Tags: pseudomerohedral twinning pseudosymmetry translational NCS X-ray crystallography translational symmetry membrane proteins bacterial amino-acid transporters MhsT research communications Source Type: research
Erwinia tasmaniensis levansucrase shows enantiomer selection for (S)-1,2,4-butanetriol
Levansucrases are biotechnologically interesting fructosyltransferases due to their potential use in the enzymatic or chemo-enzymatic synthesis of glycosides of non-natural substrates relevant to pharmaceutical applications. The structure of Erwinia tasmaniensis levansucrase in complex with (S)-1,2,4-butanetriol and its biochemical characterization suggests the possible application of short aliphatic moieties containing polyols with defined stereocentres in fructosylation biotechnology. The structural information revealed that (S)-1,2,4-butanetriol mimics the natural substrate. The preference of the protein towards a speci...
Source: Acta Crystallographica Section F - July 26, 2022 Category: Biochemistry Authors: Polsinelli, I. Salomone-Stagni, M. Benini, S. Tags: fructosyltransferases transfructosylation glycosyl hydrolases microscale thermophoresis binding assays levansucrases Erwinia tasmaniensis research communications Source Type: research
Structural study of Legionella pneumophila effector DotY (Lpg0294), a component of the Dot/Icm type IV secretion system
The bacterium Legionella pneumophila is a causative agent of Legionnaires' disease. It utilizes the Dot/Icm type IV secretion system (T4SS) to deliver over 300 effector proteins into the host cell, leading to modification of cellular processes and creating a safe environment for bacterial proliferation. Dot/Icm is a multi-subunit molecular machine. The effectors are recognized by the inner membrane-embedded coupling complex (T4CC), which then delivers them to the translocation apparatus. This T4CC subcomplex is made up of DotL, DotM, DotN, IcmS, IcmW, LvgA, DotY and DotZ, and its structure was recently determined by cryo-E...
Source: Acta Crystallographica Section F - July 4, 2022 Category: Biochemistry Authors: Chung, I.Y.W. Cygler, M. Tags: Dot/Icm secretion system type IV secretion system T4CC multi-subunit subcomplex multidomain protein domain flexibility Legionella pneumophila Legionnaires' disease research communications Source Type: research
Serial crystallography with multi-stage merging of thousands of images
KAMO and BLEND provide particularly effective tools to automatically manage the merging of large numbers of data sets from serial crystallography. The requirement for manual intervention in the process can be reduced by extending BLEND to support additional clustering options such as the use of more accurate cell distance metrics and the use of reflection-intensity correlation coefficients to infer `distances' among sets of reflections. This increases the sensitivity to differences in unit-cell parameters and allows clustering to assemble nearly complete data sets on the basis of intensity or amplitude differences. If the ...
Source: Acta Crystallographica Section F - July 1, 2022 Category: Biochemistry Authors: Soares, A.S. Yamada, Y. Jakoncic, J. McSweeney, S. Sweet, R.M. Skinner, J. Foadi, J. Fuchs, M.R. Schneider, D.K. Shi, W. Andi, B. Andrews, L.C. Bernstein, H.J. Tags: clustering serial crystallography KAMO BLEND methods communications Source Type: research
Crystal structure determination of the halogenase CtcP from Streptomyces aureofaciens
Chlortetracycline (CTC), a derivative of tetracycline (TC), is a broadly used antibiotic that inhibits the synthesis of bacterial proteins by competing with the A-site tRNA on ribosomes. A recent study showed that during the biosynthesis of CTC in Streptomyces aureofaciens, the halogenase CtcP catalyzes the final chlorination reaction and transforms TC into CTC. However, the structure of this fundamental enzyme is still lacking. Here, selenomethionine-derivatized CtcP from S. aureofaciens was overexpressed and purified and its structure was determined at 2.7 Å resolution. The structure of CtcP reveals the conserved mo...
Source: Acta Crystallographica Section F - July 1, 2022 Category: Biochemistry Authors: Yin, L. Tags: crystal structure CtcP halogenases chlortetracycline Streptomyces aureofaciens antibiotics research communications Source Type: research
