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Mechanisms of Resistance

Diversity of Substitutions within or Adjacent to Conserved Amino Acid Motifs of Penicillin-Binding Protein 2X in Cephalosporin-Resistant Streptococcus pneumoniaeIsolates

Yasuko Asahi, Yasuo Takeuchi, Kimiko Ubukata
Yasuko Asahi
Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd., Kohoku-ku, Yokohama 222-8567, and
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Yasuo Takeuchi
Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd., Kohoku-ku, Yokohama 222-8567, and
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Kimiko Ubukata
Institute of Microbial Chemistry, Shinagawa-ku, Tokyo 141-0021,
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DOI: 10.1128/AAC.43.5.1252
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  • Fig. 1.
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    Fig. 1.

    Deduced amino acid sequences of part of PBP 2X from representative strains from each group. The sequence of PSSP R6 is shown on the top line. Numbering is based on published data on the R6 strain (15). Only amino acids differing from the R6 sequence are shown. Boxes represent conserved amino acid motifs.

  • Fig. 2.
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    Fig. 2.

    Stereoview of the transpeptidase domain of PBP 2X of the high-level CTX-resistant PRSP strain 31/KU5. The structure was constructed by using the crystallographic coordinates of PBP 2X of Pares et al. (PDB entry code 1 PMD) (21). Cefditoren of oral cephalosporin was shown in the long groove as a β-lactam model. Amino acids differing from the R6 sequence are indicated by colored circles (blue, substitutions within or adjacent to conserved motifs of STMK and KTG; green, substitutions adjacent to SSN and KSG in strains classified into groups I and II; yellow, general substitutions).

Tables

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  • Table 1.

    Classification based on PBP 2X sequence differences and properties of S. pneumoniae strains

    GroupStrainSerotypeMIC (μg/ml)aAmino acid motifbPCR resultc
    PCCTXCPDXCDTRCFDNCCLS T M KA H S S N VL K S G Tpbp1apbp2b
    None1/E9150.0310.0310.1250.0160.1251–––––––––––––––−−
    2/HSB1150.0310.0310.1250.0310.1252––––V––––––––––−−
    3/YO4230.0310.0630.1250.0310.1251–––––––––––––––−−
    4/HSC2160.0310.0630.1250.0310.251–––––––––––––––−−
    5/H48190.0310.0630.1250.0310.251–––––––––––––––−−
    I6/HSB530.0080.1250.50.0310.252––––––––––––––A−−
    7/T3760.0080.1250.250.0310.252––––––––––––––A−−
    8/TJ2490.0160.1250.250.0310.251––––––––––––––A−−
    9/E1390.0160.250.50.0310.252––––––––––––––A−−
    II14/TO22190.0310.0630.250.0630.1250.5–––––L––––V––––−−
    17/SU1190.0630.1250.250.0160.251–––––L––––V––––−−
    21/H3100.0310.1250.250.0630.1251–––––L–––––––––−−
    16/MA37230.0630.1250.250.1250.251––––VL–––––––––−−
    22/H23140.0310.0630.250.0630.251––––VL–––––––––−−
    III11/KM9930.0310.1250.50.0630.250.5–A–––––––––––––−−
    15/HSC930.0310.1250.50.1250.251–A–––––––––––––−−
    18/TJ4130.0310.1250.50.0630.252–A–––––––––––––−−
    23/H6930.0310.250.250.0630.251–A–––––––––––––−−
    25/S1930.0310.1250.50.1250.251–A–––––––––––––−−
    IV34/H31610.510.5432–A––––––––V––––++
    36/Z21620.520.5864–A––––––––V––––++
    37/H281910.520.5264–A––––––––V––––++
    38/KM901920.520.5464–A––––––––V––––++
    39/Z131920.510.5464–A––––––––V––––++
    40/H292310.520.5464–A––––––––V––––++
    41/B992310.520.5464–A––––––––V––––++
    42/H0144141864–A––––––––V––––++
    27/S46230.50.510.25232–A––––––––V––––−+
    1A22/HA560.250.510.25216–A––––––––V––––−+
    V20/SHA319241643264–A––V–––––V––––++
    29/KK13323281643264–AF–––––––V––––++
    30/NG4423221621664–AF–––––––V––––++
    31/KU5231282864–AF–––––––V––––++
    32/KU8119441621664–AF–––––––V––––++
    33/AK5140.125416228–AF–––––––V––––+−
    • ↵a Susceptibilities to PC, CTX, cefpodoxime (CPDX), cefditoren (CDTR), cefdinir (CFDN), and cefaclor (CCL) were determined by previously described methods (1).

    • ↵b Only amino acid residues differing from PBP 2X conserved motif sequences of the PSSP strain R6 are shown. Conserved amino acid motifs are underlined.

    • ↵c Altered PBP genes were detected by PCR as described previously (29). +, altered; −, not altered.

  • Table 2.

    Susceptibilities to β-lactam antibiotics of S. pneumoniae transformants obtained with pbp2x DNA

    StrainaMIC (μg/ml)b
    PCCTXCPDXCDTRCFDNCCL
    Recipient, KK970.0310.0160.0310.0160.0630.5
    Transformant
     KK972X34/H310.0630.52.00.54.02.0
     KK972X40/H290.0630.52.00.54.02.0
     KK972X29/KK1330.0631.02.00.58.02.0
     KK972X32/KU810.0631.04.00.58.02.0
    • ↵a Nonencapsulated strain KK97 was transformed with PCR-amplified pbp2x from group IV and V strains as the donor DNA. Transformants were selected on plates containing 0.2 or 0.5 μg of CTX per ml. Example of transformant nomenclature, KK972X34/H31 corresponds to KK97 strain containing the pbp2x gene from strain 34/H31.

    • ↵b For abbreviations of β-lactams, see Table1, footnote a .

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Diversity of Substitutions within or Adjacent to Conserved Amino Acid Motifs of Penicillin-Binding Protein 2X in Cephalosporin-Resistant Streptococcus pneumoniaeIsolates
Yasuko Asahi, Yasuo Takeuchi, Kimiko Ubukata
Antimicrobial Agents and Chemotherapy May 1999, 43 (5) 1252-1255; DOI: 10.1128/AAC.43.5.1252

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Diversity of Substitutions within or Adjacent to Conserved Amino Acid Motifs of Penicillin-Binding Protein 2X in Cephalosporin-Resistant Streptococcus pneumoniaeIsolates
Yasuko Asahi, Yasuo Takeuchi, Kimiko Ubukata
Antimicrobial Agents and Chemotherapy May 1999, 43 (5) 1252-1255; DOI: 10.1128/AAC.43.5.1252
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KEYWORDS

Carrier Proteins
Cephalosporin Resistance
penicillin-binding proteins
Streptococcus pneumoniae

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