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Protein

Beta-lactamase

Gene

blaC

Organism
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Extended spectrum beta-lactamase (ESBL) that inactivates beta-lactam antibiotics by hydrolyzing the amide group of the beta-lactam ring. Displays high levels of penicillinase and cephalosporinase activity as well as measurable activity with carbapenems, including imipenem and meropenem. Plays a primary role in the intrinsic resistance of M.tuberculosis to beta-lactam antibiotics.5 Publications

Catalytic activityi

A beta-lactam + H2O = a substituted beta-amino acid.4 Publications

Enzyme regulationi

Is inhibited by sulbactam, tazobactam, and clavulanate. Sulbactam inhibits the enzyme competitively and reversibly with respect to nitrocefin. Tazobactam inhibits the enzyme in a time-dependent manner, but the activity of the enzyme reappears due to the slow hydrolysis of the covalently acylated enzyme. In contrast, clavulanate reacts with the enzyme quickly to form hydrolytically stable, inactive forms of the enzyme, via irreversible acylation of the catalytic serine residue. Clavulanate has potential to be used in combination with approved beta-lactam antibiotics to treat multi-drug resistant (MDR) and extremely drug resistant (XDR) strains of M.tuberculosis. Is also irreversibly inhibited by NXL104, which forms an extremely stable carbamoyl adduct with the enzyme but shows an inhibition efficiency more than 100-fold lower than that of clavulanate. Is inhibited by carbapenems, that are very poor substrates for the enzyme.5 Publications

Kineticsi

kcat is 600 min(-1) with ampicillin as substrate. kcat is 340 min(-1) with amoxicillin as substrate. kcat is 560 min(-1) with penicillin G as substrate. kcat is 2100 min(-1) with penicillin V as substrate. kcat is 690 min(-1) with piperacillin as substrate. kcat is 1070 min(-1) with cephalosporin C as substrate. kcat is 490 min(-1) with cephalotin as substrate. kcat is 490 min(-1) with cefuroxime as substrate. kcat is 3500 min(-1) with cefamandole as substrate. kcat is 48 min(-1) with cefoxitin as substrate. kcat is 2.0 min(-1) with ceftazidime as substrate. kcat is 49 min(-1) with ceftriaxone as substrate. kcat is 380 min(-1) with cefotaxime as substrate. kcat is 6680 min(-1) with nitrocefin as substrate. kcat is 1770 min(-1) with CENTA as substrate. kcat is 10 min(-1) with imipenem as substrate. kcat is 0.08 min(-1) with meropenem as substrate. Assays above performed at pH 6.4. kcat is 18.5 sec(-1) with ampicillin as substrate. kcat is 12.1 sec(-1) with cephalotin as substrate. kcat is 1.1 sec(-1) with cefoxitin as substrate. kcat is 0.2 sec(-1) with ceftazidime as substrate. kcat is 0.9 sec(-1) with meropenem as substrate. Assays above performed at pH 7.5. kcat is 0.08 min(-1) with meropenem as substrate at pH 6.5. kcat is 0.016 min(-1) with doripenem as substrate. kcat is 0.017 min(-1) with ertapenem as substrate. kcat is 0.65 min(-1) with faropenem as substrate. Assays above performed at pH 6.5.4 Publications

Manual assertion based on experiment ini

  1. KM=8 µM for ampicillin (at pH 6.4)1 Publication
  2. KM=22 µM for amoxicillin (at pH 6.4)1 Publication
  3. KM=19 µM for penicillin G (at pH 6.4)1 Publication
  4. KM=69 µM for penicillin V (at pH 6.4)1 Publication
  5. KM=59 µM for piperacillin (at pH 6.4)1 Publication
  6. KM=114 µM for cephalosporin C (at pH 6.4)1 Publication
  7. KM=152 µM for cephalotin (at pH 6.4)1 Publication
  8. KM=5100 µM for cefuroxime (at pH 6.4)1 Publication
  9. KM=184 µM for cefamandole (at pH 6.4)1 Publication
  10. KM=127 µM for cefoxitin (at pH 6.4)1 Publication
  11. KM=280 µM for ceftazidime (at pH 6.4)1 Publication
  12. KM=520 µM for ceftriaxone (at pH 6.4)1 Publication
  13. KM=5570 µM for cefotaxime (at pH 6.4)1 Publication
  14. KM=57 µM for nitrocefin (at pH 6.4)1 Publication
  15. KM=195 µM for CENTA (at pH 6.4)1 Publication
  16. KM=9.4 µM for imipenem (at pH 6.4)1 Publication
  17. KM=3.4 µM for meropenem (at pH 6.4)1 Publication
  18. KM=63 µM for ampicillin (at pH 7.5)1 Publication
  19. KM=117 µM for cephalotin (at pH 7.5)1 Publication
  20. KM=195 µM for cefoxitin (at pH 7.5)1 Publication
  21. KM=593 µM for ceftazidime (at pH 7.5)1 Publication
  22. KM=279 µM for meropenem (at pH 7.5)1 Publication
  23. KM=3.4 µM for meropenem (at pH 6.5)1 Publication
  24. KM=0.18 µM for doripenem (at pH 6.5)1 Publication
  25. KM=0.18 µM for ertapenem (at pH 6.5)1 Publication
  26. KM=55 µM for faropenem (at pH 6.5)1 Publication

    Sites

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Active sitei84Acyl-ester intermediate3 Publications1
    Sitei87Increases nucleophilicity of active site Ser2 Publications1
    Sitei117Functions as a gatekeeper residue that regulates substrate accessibility to the enzyme active site1 Publication1
    Binding sitei142Substrate3 Publications1
    Active sitei182Proton acceptor2 Publications1

    GO - Molecular functioni

    • beta-lactamase activity Source: MTBBASE
    • cephalosporinase activity Source: MTBBASE
    • penicillinase activity Source: MTBBASE

    GO - Biological processi

    • beta-lactam antibiotic catabolic process Source: MTBBASE
    • response to antibiotic Source: MTBBASE
    Complete GO annotation...

    Keywords - Molecular functioni

    Hydrolase

    Keywords - Biological processi

    Antibiotic resistance

    Enzyme and pathway databases

    BioCyciMTBH37RV:G185E-6273-MONOMER.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Beta-lactamase1 Publication (EC:3.5.2.64 Publications)
    Alternative name(s):
    Ambler class A beta-lactamase1 Publication
    Gene namesi
    Name:blaC1 Publication
    Synonyms:blaA
    Ordered Locus Names:Rv2068c
    ORF Names:MTCY49.07c
    OrganismiMycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
    Taxonomic identifieri83332 [NCBI]
    Taxonomic lineageiBacteriaActinobacteriaCorynebacterialesMycobacteriaceaeMycobacteriumMycobacterium tuberculosis complex
    Proteomesi
    • UP000001584 Componenti: Chromosome

    Organism-specific databases

    TubercuListiRv2068c.

    Subcellular locationi

    • Cell inner membrane 1 Publication
    • Periplasm 1 Publication
    • Secreted By similarity

    GO - Cellular componenti

    • extracellular region Source: MTBBASE
    • periplasmic space Source: UniProtKB-SubCell
    • plasma membrane Source: MTBBASE
    Complete GO annotation...

    Keywords - Cellular componenti

    Cell inner membrane, Cell membrane, Membrane, Periplasm, Secreted

    Pathology & Biotechi

    Biotechnological usei

    Can be used as a biomarker, which together with BlaC-specific fluorogenic substrates, allows a rapid and accurate detection of very low numbers of M.tuberculosis for the clinical diagnosis of tuberculosis in sputum and other specimens.2 Publications

    Disruption phenotypei

    Cells lacking this gene become significantly more susceptible (16- to 32-fold) to penicillins as well as third-generation cephalosporins and carbapenems. They have no detectable beta-lactamase activity.1 Publication

    Mutagenesis

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Mutagenesisi8 – 9RR → KK: No longer exported via the Tat-system. Loss of cell resistance to beta-lactam antibiotics. 1 Publication2
    Mutagenesisi87K → A: 200000-fold decrease in catalytic efficiency with cefamandole as substrate. 1 Publication1
    Mutagenesisi117I → F: Significant increase in ampicillin resistance. 2-fold and 3-fold increase in catalytic efficiency with ampicillin and nitrocefin as substrate, respectively, mainly due to an increase in substrate affinity. 1 Publication1
    Mutagenesisi142S → G: Significant reduction of catalytic activity for both nitrocefin and ampicillin. Leads to in vitro clavulanate resistance and decreased susceptibility to carbapenem inhibitors, but is still susceptible to ampicillin-clavulanate in vivo. 1 Publication1
    Mutagenesisi182E → A: Loss of catalytic activity with cefamandole as substrate. 1 Publication1
    Mutagenesisi236R → A or S: Significant reduction of catalytic activity for both nitrocefin and ampicillin. Leads to in vitro clavulanate resistance and decreased susceptibility to carbapenem inhibitors, but is still susceptible to ampicillin-clavulanate in vivo. 1 Publication1
    Mutagenesisi253T → A: Significant reduction of catalytic activity for both nitrocefin and ampicillin. Only minor impairment of the inactivation by clavulanate. Larger increase in resistance to carbapenems. 1 Publication1
    Mutagenesisi253T → S: Only minor impairment of catalytic activity with both nitrocefin and ampicillin. Still inhibited by clavulanate and carbapenems. 1 Publication1

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Signal peptidei1 – 34Tat-type signalPROSITE-ProRule annotationAdd BLAST34
    ChainiPRO_000001700535 – 307Beta-lactamaseAdd BLAST273

    Post-translational modificationi

    Exported by the Tat system. The position of the signal peptide cleavage has not been experimentally proven.1 Publication

    Proteomic databases

    PaxDbiP9WKD3.

    Expressioni

    Inductioni

    Constitutively expressed.1 Publication

    Interactioni

    Subunit structurei

    Monomer.1 Publication

    Protein-protein interaction databases

    STRINGi83332.Rv2068c.

    Structurei

    Secondary structure

    1307
    Legend: HelixTurnBeta strandPDB Structure known for this area
    Show more details
    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Helixi44 – 55Combined sources12
    Beta strandi58 – 63Combined sources6
    Beta strandi67 – 69Combined sources3
    Beta strandi72 – 75Combined sources4
    Helixi83 – 86Combined sources4
    Helixi87 – 97Combined sources11
    Helixi100 – 104Combined sources5
    Helixi111 – 113Combined sources3
    Helixi121 – 123Combined sources3
    Turni125 – 127Combined sources3
    Helixi131 – 140Combined sources10
    Helixi144 – 154Combined sources11
    Helixi158 – 160Combined sources3
    Helixi161 – 170Combined sources10
    Helixi184 – 186Combined sources3
    Beta strandi194 – 197Combined sources4
    Helixi199 – 210Combined sources12
    Beta strandi212 – 215Combined sources4
    Helixi217 – 228Combined sources12
    Turni234 – 236Combined sources3
    Helixi237 – 240Combined sources4
    Beta strandi245 – 254Combined sources10
    Turni255 – 257Combined sources3
    Beta strandi258 – 266Combined sources9
    Beta strandi272 – 280Combined sources9
    Helixi282 – 284Combined sources3
    Helixi292 – 306Combined sources15

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    PDB entryMethodResolution (Å)ChainPositionsPDBsum
    2GDNX-ray1.72A41-307[»]
    3CG5X-ray1.70A43-307[»]
    3DWZX-ray1.80A43-307[»]
    3IQAX-ray2.20A43-307[»]
    3M6BX-ray1.30A43-307[»]
    3M6HX-ray1.99A43-307[»]
    3N6IX-ray2.00A43-307[»]
    3N7WX-ray1.70A43-307[»]
    3N8LX-ray1.40A43-307[»]
    3N8RX-ray1.41A43-307[»]
    3N8SX-ray2.00A43-307[»]
    3NBLX-ray2.00A43-307[»]
    3NC8X-ray1.50A43-307[»]
    3NCKX-ray2.80A43-307[»]
    3NDEX-ray1.70A43-307[»]
    3NDGX-ray1.90A43-307[»]
    3NY4X-ray1.22A43-307[»]
    3VFFX-ray2.78A/B/C/D43-307[»]
    3VFHX-ray2.57A/B/C/D43-307[»]
    3ZHHX-ray2.85A/B/C/D32-307[»]
    4DF6X-ray2.29A43-307[»]
    4EBLX-ray2.10A/B/C/D43-307[»]
    4EBNX-ray2.85A/B/C/D43-307[»]
    4EBPX-ray2.29A/B/C/D43-307[»]
    4JLFX-ray2.10A43-307[»]
    4Q8IX-ray1.90A41-307[»]
    4QB8X-ray1.76A42-307[»]
    4QHCX-ray1.90A42-307[»]
    4X6TX-ray1.40A45-307[»]
    ProteinModelPortaliP9WKD3.
    SMRiP9WKD3.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Region

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Regioni251 – 253Substrate binding3 Publications3

    Sequence similaritiesi

    Belongs to the class-A beta-lactamase family.Curated

    Keywords - Domaini

    Signal

    Phylogenomic databases

    eggNOGiENOG4108J4B. Bacteria.
    COG2367. LUCA.
    KOiK17836.
    OMAiDEKEATY.
    PhylomeDBiP9WKD3.

    Family and domain databases

    Gene3Di3.40.710.10. 1 hit.
    InterProiIPR012338. Beta-lactam/transpept-like.
    IPR000871. Beta-lactam_class-A.
    IPR023650. Beta-lactam_class-A_AS.
    [Graphical view]
    PRINTSiPR00118. BLACTAMASEA.
    SUPFAMiSSF56601. SSF56601. 1 hit.
    PROSITEiPS00146. BETA_LACTAMASE_A. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    Sequence processingi: The displayed sequence is further processed into a mature form.

    P9WKD3-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MRNRGFGRRE LLVAMAMLVS VTGCARHASG ARPASTTLPA GADLADRFAE
    60 70 80 90 100
    LERRYDARLG VYVPATGTTA AIEYRADERF AFCSTFKAPL VAAVLHQNPL
    110 120 130 140 150
    THLDKLITYT SDDIRSISPV AQQHVQTGMT IGQLCDAAIR YSDGTAANLL
    160 170 180 190 200
    LADLGGPGGG TAAFTGYLRS LGDTVSRLDA EEPELNRDPP GDERDTTTPH
    210 220 230 240 250
    AIALVLQQLV LGNALPPDKR ALLTDWMARN TTGAKRIRAG FPADWKVIDK
    260 270 280 290 300
    TGTGDYGRAN DIAVVWSPTG VPYVVAVMSD RAGGGYDAEP REALLAEAAT

    CVAGVLA
    Length:307
    Mass (Da):32,568
    Last modified:April 16, 2014 - v1
    Checksum:i448CB2A0E05F4315
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AL123456 Genomic DNA. Translation: CCP44842.1.
    PIRiG70764.
    RefSeqiNP_216584.1. NC_000962.3.
    WP_003410677.1. NZ_KK339370.1.

    Genome annotation databases

    EnsemblBacteriaiCCP44842; CCP44842; Rv2068c.
    GeneIDi888742.
    KEGGimtu:Rv2068c.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AL123456 Genomic DNA. Translation: CCP44842.1.
    PIRiG70764.
    RefSeqiNP_216584.1. NC_000962.3.
    WP_003410677.1. NZ_KK339370.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    PDB entryMethodResolution (Å)ChainPositionsPDBsum
    2GDNX-ray1.72A41-307[»]
    3CG5X-ray1.70A43-307[»]
    3DWZX-ray1.80A43-307[»]
    3IQAX-ray2.20A43-307[»]
    3M6BX-ray1.30A43-307[»]
    3M6HX-ray1.99A43-307[»]
    3N6IX-ray2.00A43-307[»]
    3N7WX-ray1.70A43-307[»]
    3N8LX-ray1.40A43-307[»]
    3N8RX-ray1.41A43-307[»]
    3N8SX-ray2.00A43-307[»]
    3NBLX-ray2.00A43-307[»]
    3NC8X-ray1.50A43-307[»]
    3NCKX-ray2.80A43-307[»]
    3NDEX-ray1.70A43-307[»]
    3NDGX-ray1.90A43-307[»]
    3NY4X-ray1.22A43-307[»]
    3VFFX-ray2.78A/B/C/D43-307[»]
    3VFHX-ray2.57A/B/C/D43-307[»]
    3ZHHX-ray2.85A/B/C/D32-307[»]
    4DF6X-ray2.29A43-307[»]
    4EBLX-ray2.10A/B/C/D43-307[»]
    4EBNX-ray2.85A/B/C/D43-307[»]
    4EBPX-ray2.29A/B/C/D43-307[»]
    4JLFX-ray2.10A43-307[»]
    4Q8IX-ray1.90A41-307[»]
    4QB8X-ray1.76A42-307[»]
    4QHCX-ray1.90A42-307[»]
    4X6TX-ray1.40A45-307[»]
    ProteinModelPortaliP9WKD3.
    SMRiP9WKD3.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    STRINGi83332.Rv2068c.

    Proteomic databases

    PaxDbiP9WKD3.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiCCP44842; CCP44842; Rv2068c.
    GeneIDi888742.
    KEGGimtu:Rv2068c.

    Organism-specific databases

    TubercuListiRv2068c.

    Phylogenomic databases

    eggNOGiENOG4108J4B. Bacteria.
    COG2367. LUCA.
    KOiK17836.
    OMAiDEKEATY.
    PhylomeDBiP9WKD3.

    Enzyme and pathway databases

    BioCyciMTBH37RV:G185E-6273-MONOMER.

    Family and domain databases

    Gene3Di3.40.710.10. 1 hit.
    InterProiIPR012338. Beta-lactam/transpept-like.
    IPR000871. Beta-lactam_class-A.
    IPR023650. Beta-lactam_class-A_AS.
    [Graphical view]
    PRINTSiPR00118. BLACTAMASEA.
    SUPFAMiSSF56601. SSF56601. 1 hit.
    PROSITEiPS00146. BETA_LACTAMASE_A. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Entry informationi

    Entry nameiBLAC_MYCTU
    AccessioniPrimary (citable) accession number: P9WKD3
    Secondary accession number(s): L0T8I9
    , P0A5I6, P0C5C1, Q10670
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: April 16, 2014
    Last sequence update: April 16, 2014
    Last modified: November 2, 2016
    This is version 20 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure, Complete proteome, Reference proteome

    Documents

    1. Mycobacterium tuberculosis strains ATCC 25618 / H37Rv and CDC 1551 / Oshkosh
      Mycobacterium tuberculosis strains ATCC 25618 / H37Rv and CDC 1551 / Oshkosh: entries and gene names
    2. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    3. SIMILARITY comments
      Index of protein domains and families

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.