Structural characterization of Porphyromonas gingivalis enoyl-ACP reductase II (FabK)

Kirk Hevener, Bernard D. Santarsiero, Hyun Lee, Jesse A. Jones, Teuta Boci, Michael E. Johnson, Shahila Mehboob

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Enoyl-acyl carrier protein (ACP) reductase II (FabK) is a critical rate-limiting enzyme in the bacterial type II fatty-acid synthesis (FAS II) pathway. FAS II pathway enzymes are markedly disparate from their mammalian analogs in the FAS I pathway in both structure and mechanism. Enzymes involved in bacterial fatty-acid synthesis represent viable drug targets for Gram-negative pathogens, and historical precedent exists for targeting them in the treatment of diseases of the oral cavity. The Gram-negative organism Porphyromonas gingivalis represents a key causative agent of the costly and highly prevalent disease known as chronic periodontitis, and exclusively expresses FabK as its enoyl reductase enzyme in the FAS-II pathway. Together, these characteristics distinguish P. gingivalis FabK (PgFabK) as an attractive and novel narrow-spectrum antibacterial target candidate. PgFabK is a flavoenzyme that is dependent on FMN and NADPH as cofactors for the enzymatic reaction, which reduces the enoyl substrate via a ping-pong mechanism. Here, the structure of the PgFabK enzyme as determined using X-ray crystallography is reported to 1.9 Å resolution with endogenous FMN fully resolved and the NADPH cofactor partially resolved. PgFabK possesses a TIM-barrel motif, and all flexible loops are visible. The determined structure has allowed insight into the structural basis for the NADPH dependence observed in PgFabK and the role of a monovalent cation that has been observed in previous studies to be stringently required for FabK activity. The PgFabK structure and the insights gleaned from its analysis will facilitate structure-based drug-discovery efforts towards the prevention and treatment of P. gingivalis infection.The structure of Porphyromonas gingivalis enoyl-ACP reductase II (FabK) was determined via X-ray crystallography to 1.9 Å resolution with all flexible loops visible, FMN fully resolved and the NADPH cofactor partially resolved. Characterization includes the structural basis of NADPH dependence and the role of a required monovalent cation.

Original languageEnglish (US)
Pages (from-to)105-112
Number of pages8
JournalActa Crystallographica Section F: Structural Biology Communications
Volume74
Issue number2
DOIs
StatePublished - Feb 1 2018

Fingerprint

Acyl Carrier Protein
Porphyromonas gingivalis
NADP
enzymes
Oxidoreductases
Flavin Mononucleotide
proteins
Enzymes
Monovalent Cations
X ray crystallography
fatty acids
crystallography
drugs
Fatty Acids
cations
X Ray Crystallography
pathogens
Pathogens
infectious diseases
synthesis

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

Cite this

Structural characterization of Porphyromonas gingivalis enoyl-ACP reductase II (FabK). / Hevener, Kirk; Santarsiero, Bernard D.; Lee, Hyun; Jones, Jesse A.; Boci, Teuta; Johnson, Michael E.; Mehboob, Shahila.

In: Acta Crystallographica Section F: Structural Biology Communications, Vol. 74, No. 2, 01.02.2018, p. 105-112.

Research output: Contribution to journalArticle

Hevener, Kirk ; Santarsiero, Bernard D. ; Lee, Hyun ; Jones, Jesse A. ; Boci, Teuta ; Johnson, Michael E. ; Mehboob, Shahila. / Structural characterization of Porphyromonas gingivalis enoyl-ACP reductase II (FabK). In: Acta Crystallographica Section F: Structural Biology Communications. 2018 ; Vol. 74, No. 2. pp. 105-112.
@article{e7cf7dc0125144b0a391e139a42e0a09,
title = "Structural characterization of Porphyromonas gingivalis enoyl-ACP reductase II (FabK)",
abstract = "Enoyl-acyl carrier protein (ACP) reductase II (FabK) is a critical rate-limiting enzyme in the bacterial type II fatty-acid synthesis (FAS II) pathway. FAS II pathway enzymes are markedly disparate from their mammalian analogs in the FAS I pathway in both structure and mechanism. Enzymes involved in bacterial fatty-acid synthesis represent viable drug targets for Gram-negative pathogens, and historical precedent exists for targeting them in the treatment of diseases of the oral cavity. The Gram-negative organism Porphyromonas gingivalis represents a key causative agent of the costly and highly prevalent disease known as chronic periodontitis, and exclusively expresses FabK as its enoyl reductase enzyme in the FAS-II pathway. Together, these characteristics distinguish P. gingivalis FabK (PgFabK) as an attractive and novel narrow-spectrum antibacterial target candidate. PgFabK is a flavoenzyme that is dependent on FMN and NADPH as cofactors for the enzymatic reaction, which reduces the enoyl substrate via a ping-pong mechanism. Here, the structure of the PgFabK enzyme as determined using X-ray crystallography is reported to 1.9 {\AA} resolution with endogenous FMN fully resolved and the NADPH cofactor partially resolved. PgFabK possesses a TIM-barrel motif, and all flexible loops are visible. The determined structure has allowed insight into the structural basis for the NADPH dependence observed in PgFabK and the role of a monovalent cation that has been observed in previous studies to be stringently required for FabK activity. The PgFabK structure and the insights gleaned from its analysis will facilitate structure-based drug-discovery efforts towards the prevention and treatment of P. gingivalis infection.The structure of Porphyromonas gingivalis enoyl-ACP reductase II (FabK) was determined via X-ray crystallography to 1.9 {\AA} resolution with all flexible loops visible, FMN fully resolved and the NADPH cofactor partially resolved. Characterization includes the structural basis of NADPH dependence and the role of a required monovalent cation.",
author = "Kirk Hevener and Santarsiero, {Bernard D.} and Hyun Lee and Jones, {Jesse A.} and Teuta Boci and Johnson, {Michael E.} and Shahila Mehboob",
year = "2018",
month = "2",
day = "1",
doi = "10.1107/S2053230X18000262",
language = "English (US)",
volume = "74",
pages = "105--112",
journal = "Acta Crystallographica Section F:Structural Biology Communications",
issn = "1744-3091",
publisher = "John Wiley and Sons Ltd",
number = "2",

}

TY - JOUR

T1 - Structural characterization of Porphyromonas gingivalis enoyl-ACP reductase II (FabK)

AU - Hevener, Kirk

AU - Santarsiero, Bernard D.

AU - Lee, Hyun

AU - Jones, Jesse A.

AU - Boci, Teuta

AU - Johnson, Michael E.

AU - Mehboob, Shahila

PY - 2018/2/1

Y1 - 2018/2/1

N2 - Enoyl-acyl carrier protein (ACP) reductase II (FabK) is a critical rate-limiting enzyme in the bacterial type II fatty-acid synthesis (FAS II) pathway. FAS II pathway enzymes are markedly disparate from their mammalian analogs in the FAS I pathway in both structure and mechanism. Enzymes involved in bacterial fatty-acid synthesis represent viable drug targets for Gram-negative pathogens, and historical precedent exists for targeting them in the treatment of diseases of the oral cavity. The Gram-negative organism Porphyromonas gingivalis represents a key causative agent of the costly and highly prevalent disease known as chronic periodontitis, and exclusively expresses FabK as its enoyl reductase enzyme in the FAS-II pathway. Together, these characteristics distinguish P. gingivalis FabK (PgFabK) as an attractive and novel narrow-spectrum antibacterial target candidate. PgFabK is a flavoenzyme that is dependent on FMN and NADPH as cofactors for the enzymatic reaction, which reduces the enoyl substrate via a ping-pong mechanism. Here, the structure of the PgFabK enzyme as determined using X-ray crystallography is reported to 1.9 Å resolution with endogenous FMN fully resolved and the NADPH cofactor partially resolved. PgFabK possesses a TIM-barrel motif, and all flexible loops are visible. The determined structure has allowed insight into the structural basis for the NADPH dependence observed in PgFabK and the role of a monovalent cation that has been observed in previous studies to be stringently required for FabK activity. The PgFabK structure and the insights gleaned from its analysis will facilitate structure-based drug-discovery efforts towards the prevention and treatment of P. gingivalis infection.The structure of Porphyromonas gingivalis enoyl-ACP reductase II (FabK) was determined via X-ray crystallography to 1.9 Å resolution with all flexible loops visible, FMN fully resolved and the NADPH cofactor partially resolved. Characterization includes the structural basis of NADPH dependence and the role of a required monovalent cation.

AB - Enoyl-acyl carrier protein (ACP) reductase II (FabK) is a critical rate-limiting enzyme in the bacterial type II fatty-acid synthesis (FAS II) pathway. FAS II pathway enzymes are markedly disparate from their mammalian analogs in the FAS I pathway in both structure and mechanism. Enzymes involved in bacterial fatty-acid synthesis represent viable drug targets for Gram-negative pathogens, and historical precedent exists for targeting them in the treatment of diseases of the oral cavity. The Gram-negative organism Porphyromonas gingivalis represents a key causative agent of the costly and highly prevalent disease known as chronic periodontitis, and exclusively expresses FabK as its enoyl reductase enzyme in the FAS-II pathway. Together, these characteristics distinguish P. gingivalis FabK (PgFabK) as an attractive and novel narrow-spectrum antibacterial target candidate. PgFabK is a flavoenzyme that is dependent on FMN and NADPH as cofactors for the enzymatic reaction, which reduces the enoyl substrate via a ping-pong mechanism. Here, the structure of the PgFabK enzyme as determined using X-ray crystallography is reported to 1.9 Å resolution with endogenous FMN fully resolved and the NADPH cofactor partially resolved. PgFabK possesses a TIM-barrel motif, and all flexible loops are visible. The determined structure has allowed insight into the structural basis for the NADPH dependence observed in PgFabK and the role of a monovalent cation that has been observed in previous studies to be stringently required for FabK activity. The PgFabK structure and the insights gleaned from its analysis will facilitate structure-based drug-discovery efforts towards the prevention and treatment of P. gingivalis infection.The structure of Porphyromonas gingivalis enoyl-ACP reductase II (FabK) was determined via X-ray crystallography to 1.9 Å resolution with all flexible loops visible, FMN fully resolved and the NADPH cofactor partially resolved. Characterization includes the structural basis of NADPH dependence and the role of a required monovalent cation.

UR - http://www.scopus.com/inward/record.url?scp=85041409273&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85041409273&partnerID=8YFLogxK

U2 - 10.1107/S2053230X18000262

DO - 10.1107/S2053230X18000262

M3 - Article

VL - 74

SP - 105

EP - 112

JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

SN - 1744-3091

IS - 2

ER -