3.4.21.53: Endopeptidase La

670826

AAA+ alpha domain, containing four alpha helices and two parallel strands

670039

carboxy-terminal domain crystallized

682821

digestion of lonA by alpha-chymotrypsin yields a stable fragment consisting of residues 491-584 crystallized. Crystal structure of the proteolytic domain of lonA (residues 585-784) elucidates a unique fold, P31 space group

682726

inactive Lon-S679A P-domain successfully crystallized by the hanging drop vapor diffusion method

651835

investigation of the mode of peptide interaction with the proteolytically inactive Lon mutant S679A in the absence and presence of ADP or AMPPNP shows that the binding interaction between protein and peptide varies with the nucleotide bound to the enzyme

696457

N-terminal residues 1-245 comprising most of the N-terminal domain of the enzyme, as selenomethionine derivative to 2.6 A resolution. The molecule consists of two compact subdomains and a very long C-terminal alpha-helix. The structure of the first subdomain, residues 1-117, which consists mostly of beta-strands, is similar to that of the shorter fragment, whereas the second subdomain is almost entirely helical. The fold and spatial relationship of the two subdomains, with the exception of the C-terminal helix, closely resemble the structure of BPP1347, a 203-amino-acid protein of unknown function from Bordetella parapertussis

P0A9M0

717026

purified enzyme mutant S855A in complex with non-hydrolyzable ATP analogue AMP-PNP and with ADP, X-ray diffraction structure determination and analysis at resolutions of 15 A and 21 A, respectively, fitting of X-rays structures and cryo-electron microscopy structures, overview

Homo sapiens

P36776

755444

structure of the proteolytic domain of human mitochondrial Lon at 2 A resolution. There are six protomers in the asymmetric unit, four arranged as two dimers. The intersubunit interactions within the two dimers are similar to those between adjacent subunits of the hexameric ring of Escherichia coli Lon. The active site contains a 310 helix attached to the N-terminal end of alpha-helix 2, which leads to the insertion of Asp852 into the active site. Structural considerations make it likely that this conformation is proteolytically inactive. A mechanism relating the formation of Lon oligomers with a conformational shift in the active site region coupled to a movement of a loop in the oligomer interface may convert the proteolytically inactive form seen here to the active one in the Escherichia coli hexamer

Homo sapiens

P36776

710500

purified mutant core-E423Q in complex with Mg2+, X-ray diffraction structure determination and analysis

755562

purified recombinant full-length wild-type enzyme LonA in complex with bortezomib and Mg2+, and in apoform with Mg2+, hexameric and dodecameric enzyme crystals, AAAP-Mg2+-bortezomib complex by hanging drop vapor diffusion method, for the apoenzyme-Mg2+ crystals, 400 nl of 8 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 100 mM NaCl, and 10 mM MgCl2, is mixed with 400 nl of well solution containing 20% PEG 3000, 0.1 M HEPES, pH 7.5, and 0.2 M NaCl, for the complex crystals, mixing of 0.001 ml of protein-inhibitor solution with 0.001 ml of well solution containing 0.2 M sodium citrate, pH 6.5, and 10% PEG 3350, 22°C, X-ray diffraction structure determination and analysis at 1.85 A and 3.05 A resolution, respectively, molecular replacement

A0A059VAZ3

755563

purified recombinant selenomethionine-labeled N-terminal domain mutant L91M/L188M/I359M , mixing of 0.001 ml of 8 mg/ml protein in 10 mM HEPES, pH 7.5, 100 mM NaCl, 10% v/v glycerol, 1 mM DTT, with 0.001 ml of well solution containing consisting of 400 mM lithium sulfate, 10 mM nickel chloride, 100 mM Tris-HCl, pH 8.4, 22°C, 7 days to 3 months, Selected crystals are further dehydrated for 3 days in sitting drops equilibrated against 0.7-1.2 M lithium sulfate, X-ray diffraction structure determination and analysis at 3.1-3.4 A resolution, modelling

C9DRU9

731056

purified selenomethionine-labeled wild-type and mutant L91M/I359M enzymes alone and in complexes with inhibitors bortezomib, MG262, and clasto-lactacystin beta-lactone, sitting drop vapour diffusion technique, using a reservoir consisting of 10% 2-propanol, 100 mM dipotassium phosphate, 100 mM sodium citrate, pH 4.6, 22°C, X-ray diffraction structure determination and analysis at 1.9-2.1 A resolution

Methanocaldococcus jannaschii

731052

contains two transmembrane helices within its N-terminal domain

669297

crystal structure determination at 2.0 A resolution

B6YU74

752803, 753673

deletion mutant lacking the putative membrane-anchoring region, residues 134-170, and introduction of mutations S523A and K566A, to 2.0 A resolution. The structure is a three-tiered hexagonal cylinder with a large sequestered chamber accessible through an axial channel. Conserved loops extending from the ATPase domain combine with an insertion domain containing the membrane anchor to form an apical domain that serves as a gate governing substrate access to an internal unfolding and degradation chamber. Alternating ATPase domains are in tight- and weak-binding nucleotide states with different domain orientations and intersubunit contacts

B6YU74

717537

purified isolated AAA+ module of LonB protease, mixing of 0.001 ml of 12 mg/ml protein solution in 10 mM Tris-HCl, pH 7.5, 100 mM NaCl, and 1 mM DTT, with 0.001 ml of reservoir solution containing 0.2 M potassium chloride, 0.01 M magnesium acetate, 0.05 M tri-sodium citrate dihydrate, pH 4.5, 12% PEG 4000, and 5% w/v n-dodecyl-beta-D-maltoside, 22°C, X-ray diffraction structure determination and analysis at 2.03 A resolution

754406

to 2.0 A resolution, space group P63