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- PDB-9t7m: Crystallographic structure of Clostridioides difficile CspA prote... -

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Basic information

Entry
Database: PDB / ID: 9t7m
TitleCrystallographic structure of Clostridioides difficile CspA protein in complex with Taurocholate at 2.9 angstrom resolution
ComponentsSubtilisin-like serine germination related protease
KeywordsSIGNALING PROTEIN / Pseudo Protease / Clostridium / spore germination
Function / homology
Function and homology information


subtilisin / serine-type endopeptidase activity / proteolysis / :
Similarity search - Function
Csp protease B, prodomain / Csp protease B prodomain / CspA-like domain / : / Serine proteases, subtilase family, serine active site. / Peptidase S8, subtilisin, Ser-active site / Peptidase S8, subtilisin-related / Serine proteases, subtilase domain profile. / Peptidase S8/S53 domain superfamily / Subtilase family / Peptidase S8/S53 domain
Similarity search - Domain/homology
TAUROCHOLIC ACID / Subtilisin-like serine germination related protease
Similarity search - Component
Biological speciesClostridioides difficile (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.9 Å
AuthorsAlcorlo, M. / Mayor-Luna, C. / Hermoso, J.
Funding support Spain, 1items
OrganizationGrant numberCountry
Spanish Ministry of Science, Innovation, and UniversitiesPID2023-153118OB-I00 Spain
CitationJournal: To be published
Title: Crystallographic structure of Clostridioides difficile CspA protein in complex with Taurocholate at 2.9 angstrom resolution
Authors: Alcorlo, M. / Mayor-Luna, C. / Hermoso, J.
History
DepositionNov 11, 2025Deposition site: PDBE / Processing site: PDBE
Revision 1.0Jun 17, 2026Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
E: Subtilisin-like serine germination related protease
A: Subtilisin-like serine germination related protease
B: Subtilisin-like serine germination related protease
C: Subtilisin-like serine germination related protease
D: Subtilisin-like serine germination related protease
F: Subtilisin-like serine germination related protease
hetero molecules


Theoretical massNumber of molelcules
Total (without water)364,17314
Polymers360,0486
Non-polymers4,1268
Water00
1
E: Subtilisin-like serine germination related protease
C: Subtilisin-like serine germination related protease
hetero molecules


Theoretical massNumber of molelcules
Total (without water)121,5635
Polymers120,0162
Non-polymers1,5473
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area6080 Å2
ΔGint-10 kcal/mol
Surface area38170 Å2
MethodPISA
2
A: Subtilisin-like serine germination related protease
hetero molecules


Theoretical massNumber of molelcules
Total (without water)60,5242
Polymers60,0081
Non-polymers5161
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area980 Å2
ΔGint-0 kcal/mol
Surface area20890 Å2
MethodPISA
3
B: Subtilisin-like serine germination related protease
D: Subtilisin-like serine germination related protease
hetero molecules


Theoretical massNumber of molelcules
Total (without water)121,0474
Polymers120,0162
Non-polymers1,0312
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5600 Å2
ΔGint-11 kcal/mol
Surface area38510 Å2
MethodPISA
4
F: Subtilisin-like serine germination related protease
hetero molecules


Theoretical massNumber of molelcules
Total (without water)61,0393
Polymers60,0081
Non-polymers1,0312
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area1840 Å2
ΔGint3 kcal/mol
Surface area21210 Å2
MethodPISA
Unit cell
Length a, b, c (Å)141.239, 158.629, 188.364
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number18
Space group name H-MP22121

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Components

#1: Protein
Subtilisin-like serine germination related protease


Mass: 60007.977 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Clostridioides difficile (bacteria) / Gene: cspBA, SAMEA1402399_00499
Production host: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria)
References: UniProt: A0AB74Q767, subtilisin
#2: Chemical
ChemComp-TCH / TAUROCHOLIC ACID


Mass: 515.703 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C26H45NO7S / Feature type: SUBJECT OF INVESTIGATION
Has ligand of interestY
Has protein modificationN

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 2.94 Å3/Da / Density % sol: 58.11 %
Crystal growTemperature: 291 K / Method: vapor diffusion, sitting drop / Details: 0.2 M Na Sulfate and 20 % PEG 3350

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: ALBA / Beamline: XALOC / Wavelength: 0.97918 Å
DetectorType: DECTRIS PILATUS3 S 6M / Detector: PIXEL / Date: Oct 17, 2025
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97918 Å / Relative weight: 1
ReflectionResolution: 2.9→49.52 Å / Num. obs: 94280 / % possible obs: 100 % / Redundancy: 13.4 % / CC1/2: 0.996 / Net I/σ(I): 14.2
Reflection shellResolution: 2.9→2.95 Å / Num. unique obs: 4617 / CC1/2: 0.812

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Processing

Software
NameVersionClassification
PHENIX(1.21.2_5419: ???)refinement
PDB_EXTRACTdata extraction
XDSdata reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.9→28.87 Å / SU ML: 0.33 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 23.35 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2406 4707 5 %
Rwork0.2051 --
obs0.2069 94107 99.98 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.1 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.9→28.87 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms25211 0 280 0 25491
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00326013
X-RAY DIFFRACTIONf_angle_d0.53535403
X-RAY DIFFRACTIONf_dihedral_angle_d16.0989925
X-RAY DIFFRACTIONf_chiral_restr0.0454035
X-RAY DIFFRACTIONf_plane_restr0.0054556
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.9-2.930.36911590.31112921X-RAY DIFFRACTION100
2.93-2.970.35481600.29052921X-RAY DIFFRACTION100
2.97-30.31241500.29642986X-RAY DIFFRACTION100
3-3.040.30981720.27112914X-RAY DIFFRACTION100
3.04-3.080.33441560.26832961X-RAY DIFFRACTION100
3.08-3.120.281760.2612944X-RAY DIFFRACTION100
3.12-3.170.30741450.25092925X-RAY DIFFRACTION100
3.17-3.220.27681680.25532949X-RAY DIFFRACTION100
3.22-3.270.30481500.25032985X-RAY DIFFRACTION100
3.27-3.320.30071470.25572943X-RAY DIFFRACTION100
3.32-3.380.27651560.23242976X-RAY DIFFRACTION100
3.38-3.440.30851570.24072931X-RAY DIFFRACTION100
3.44-3.50.25441570.22782958X-RAY DIFFRACTION100
3.5-3.570.24641510.22692957X-RAY DIFFRACTION100
3.57-3.650.25431610.22232958X-RAY DIFFRACTION100
3.65-3.740.27431680.21772967X-RAY DIFFRACTION100
3.74-3.830.24821670.21562956X-RAY DIFFRACTION100
3.83-3.930.26061350.20553003X-RAY DIFFRACTION100
3.93-4.050.23481510.18942966X-RAY DIFFRACTION100
4.05-4.180.22021590.19132975X-RAY DIFFRACTION100
4.18-4.330.2181470.17372971X-RAY DIFFRACTION100
4.33-4.50.17981290.16253026X-RAY DIFFRACTION100
4.5-4.710.20691700.15842978X-RAY DIFFRACTION100
4.71-4.950.20161840.16022953X-RAY DIFFRACTION100
4.95-5.260.20231530.17723019X-RAY DIFFRACTION100
5.26-5.660.19521540.19083024X-RAY DIFFRACTION100
5.66-6.230.25881630.20842997X-RAY DIFFRACTION100
6.23-7.120.24151560.20663058X-RAY DIFFRACTION100
7.12-8.920.18261460.16933095X-RAY DIFFRACTION100
8.92-28.870.17051600.16223183X-RAY DIFFRACTION99
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
12.6146-0.39990.18571.46640.40421.7598-0.0464-0.20010.2510.11920.0083-0.1717-0.09650.08740.02860.30490.0159-0.04650.20830.0320.1865-29.1404-6.744529.9416
21.6361-0.0928-1.20250.9075-0.50932.7726-0.0479-0.48540.26410.37410.1583-0.3817-0.09671.0205-0.17620.3365-0.0043-0.06870.4274-0.03040.3783-18.0233-15.584836.1815
31.46930.10490.52070.8257-0.0520.84150.01280.108-0.05970.0179-0.0886-0.30990.02140.27530.06350.28720.0315-0.03140.30430.06160.3245-10.0447-17.489827.5849
41.3883-0.51360.35981.9613-0.66651.61-0.0472-0.09230.0136-0.05960.07990.04540.0327-0.0209-0.05670.24860.0032-0.00520.1986-0.04620.2034-44.2732-27.0489-4.4953
51.5292-0.2828-0.16130.89260.04730.5278-0.04830.0402-0.0177-0.20730.0955-0.21420.02320.1042-0.03830.3442-0.01410.03610.2126-0.0420.2179-30.5049-32.4261-17.9837
62.7668-0.09760.11180.6729-0.36122.51180.05740.11280.0214-0.1823-0.0644-0.52370.20340.68090.07220.4436-0.01810.06160.68030.10330.4747-54.783-18.113441.8566
71.027-0.14160.16061.63130.07231.6237-0.01050.33160.3603-0.2177-0.09850.0827-0.286-0.04740.08590.45020.05050.00410.4740.14950.4287-82.5922-8.810339.254
81.1057-0.6344-0.17572.15861.22021.848-0.02220.38240.10030.5620.1544-0.60620.13350.5251-0.07180.3472-0.0037-0.06240.4040.02810.3874-77.1296-19.12652.5037
90.67360.0613-0.14991.67870.08311.98650.10420.4209-0.3391-0.2806-0.0719-0.21660.33910.181-0.02380.42940.14740.00370.5891-0.09470.416-65.8263-44.13944.6227
101.1824-0.72810.11571.42110.06171.05320.0220.2497-0.00320.1271-0.07820.1540.031-0.01680.0550.3026-0.00540.04960.3750.01540.309-83.978-25.321750.901
114.4793-0.10350.26280.73530.88313.2535-0.2612-0.16480.41460.0868-0.19310.33030.3217-0.17360.29640.4122-0.00450.00670.43560.07770.5489-28.2293-15.856567.6769
122.33920.29680.22651.3524-0.20821.9537-0.0703-0.13830.26540.24140.0382-0.217-0.20910.03960.00840.42670.0018-0.18140.28410.02050.51140.547-8.8270.4459
130.62290.9329-0.27651.7586-0.28050.73840.1845-0.1338-0.0893-0.37560.04940.4690.051-0.1567-0.22450.4580.0344-0.14340.33540.03570.558-4.6066-17.03655.9966
140.5711-0.08780.03281.6898-0.41881.7230.1456-0.2914-0.40540.10820.0680.11530.2741-0.3506-0.1990.5407-0.0929-0.18320.45330.16810.5716-18.2546-40.762759.593
151.34960.69920.06981.0643-0.09881.37440.0071-0.0118-0.0856-0.02240.0363-0.20950.07240.2071-0.02060.31120.0157-0.11480.32480.03070.51764.7091-20.390655.9439
162.52190.26390.13332.0952-0.75781.4049-0.00120.64461.5832-0.50860.2292-0.135-0.6357-0.2784-0.25010.7162-0.06170.0270.63510.14680.9449-67.78052.391274.145
172.2133-0.2852-0.25871.71280.2331.8130.0247-0.34830.29550.3191-0.0279-0.2577-0.07980.52640.01650.4165-0.0847-0.04720.5726-0.04730.372-48.6375-20.455881.9854
181.8346-0.304-1.76140.6597-0.2633.30710.06390.53280.15170.081-0.0934-0.2548-0.588-0.5238-0.07680.3924-0.0186-0.01910.49020.07860.3646-64.4093-22.840772.7709
191.2019-0.3012-0.15081.60670.39351.6224-0.0557-0.3321-0.06360.5405-0.07280.13730.2189-0.08770.05740.4977-0.06270.09420.3871-0.04440.3436-86.0759-19.15886.6677
200.5808-0.1077-0.87260.2175-0.00192.6526-0.03490.00990.35780.2750.01470.2521-0.3292-0.40160.01530.61410.04360.11560.4867-0.08310.4773-88.8173-9.838982.6101
212.0181-0.6129-0.18810.72830.24121.4342-0.0052-0.2571-0.07260.2804-0.03340.0956-0.0010.09210.01260.453-0.04410.03920.29580.03230.3132-73.1771-25.267184.031
222.71630.0380.30021.63-0.08731.0081-0.01470.0809-0.47620.1088-0.1236-0.19240.13550.41050.10690.39180.02040.0510.47410.02370.3139-56.4773-36.052569.1273
231.3397-0.9284-0.25240.96950.05511.3161-0.0661-1.05080.43590.54490.23560.2159-0.0546-0.01750.03050.901-0.0387-0.11391.2402-0.08530.6426-29.344848.391189.6568
240.71080.20350.62441.6448-0.38760.77690.092-0.3729-1.0431-0.0628-0.0316-0.40970.24130.091-0.08340.5196-0.0432-0.16540.5290.15740.8821-4.96827.216969.0223
250.79840.04520.15860.9922-0.26440.74210.2497-0.8494-0.48870.3809-0.04-0.44890.3889-0.1242-0.1720.6448-0.1549-0.31330.84570.27610.7954-5.53829.394382.6373
261.4647-0.23990.09610.7964-0.25190.59090.2324-0.5508-0.16630.1553-0.0478-0.13580.0619-0.1729-0.1690.4903-0.102-0.09890.5030.09190.47-15.880741.447973.0654
270.86340.0504-0.0180.75940.11390.91770.1987-0.4419-0.32490.0692-0.01510.11030.2441-0.2073-0.07430.5475-0.1185-0.12740.69990.17070.4456-43.756435.127466.5704
281.97490.83430.19170.9014-0.04750.70170.0218-0.9965-0.1630.61170.25110.64740.0919-1.2543-0.15530.6239-0.23690.00561.17390.24120.6898-50.54536.356477.0373
290.81060.1339-0.06190.3714-0.10081.09840.1665-0.5169-0.43010.593-0.0111-0.1051-0.0619-0.2188-0.18770.5615-0.1352-0.12650.74760.20760.5108-40.907235.947569.1063
301.7315-0.0829-0.01680.35510.08821.36880.2033-0.0831-0.51170.0198-0.0257-0.2110.2785-0.1869-0.17720.4426-0.0799-0.15550.38990.10760.583-15.482932.958863.2668
311.91050.04170.51621.01450.08440.9983-0.1190.1817-0.511-0.10190.2817-0.21930.18620.1092-0.12790.4274-0.066-0.09230.459-0.05860.5515-9.96938.047152.8193
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'E' and (resid 587 through 833 )
2X-RAY DIFFRACTION2chain 'E' and (resid 834 through 862 )
3X-RAY DIFFRACTION3chain 'E' and (resid 863 through 1132 )
4X-RAY DIFFRACTION4chain 'A' and (resid 587 through 797 )
5X-RAY DIFFRACTION5chain 'A' and (resid 798 through 1134 )
6X-RAY DIFFRACTION6chain 'B' and (resid 587 through 635 )
7X-RAY DIFFRACTION7chain 'B' and (resid 636 through 833 )
8X-RAY DIFFRACTION8chain 'B' and (resid 834 through 862 )
9X-RAY DIFFRACTION9chain 'B' and (resid 863 through 939 )
10X-RAY DIFFRACTION10chain 'B' and (resid 940 through 1132 )
11X-RAY DIFFRACTION11chain 'C' and (resid 587 through 635 )
12X-RAY DIFFRACTION12chain 'C' and (resid 636 through 833 )
13X-RAY DIFFRACTION13chain 'C' and (resid 834 through 862 )
14X-RAY DIFFRACTION14chain 'C' and (resid 863 through 970 )
15X-RAY DIFFRACTION15chain 'C' and (resid 971 through 1133 )
16X-RAY DIFFRACTION16chain 'D' and (resid 587 through 645 )
17X-RAY DIFFRACTION17chain 'D' and (resid 646 through 833 )
18X-RAY DIFFRACTION18chain 'D' and (resid 834 through 862 )
19X-RAY DIFFRACTION19chain 'D' and (resid 863 through 894 )
20X-RAY DIFFRACTION20chain 'D' and (resid 895 through 947 )
21X-RAY DIFFRACTION21chain 'D' and (resid 948 through 1062 )
22X-RAY DIFFRACTION22chain 'D' and (resid 1063 through 1133 )
23X-RAY DIFFRACTION23chain 'F' and (resid 587 through 645 )
24X-RAY DIFFRACTION24chain 'F' and (resid 646 through 703 )
25X-RAY DIFFRACTION25chain 'F' and (resid 704 through 797 )
26X-RAY DIFFRACTION26chain 'F' and (resid 798 through 862 )
27X-RAY DIFFRACTION27chain 'F' and (resid 863 through 905 )
28X-RAY DIFFRACTION28chain 'F' and (resid 906 through 939 )
29X-RAY DIFFRACTION29chain 'F' and (resid 940 through 989 )
30X-RAY DIFFRACTION30chain 'F' and (resid 990 through 1092 )
31X-RAY DIFFRACTION31chain 'F' and (resid 1093 through 1134 )

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