Structure and function of Mycobacterium smegmatis 7-keto-8-aminopelargonic acid (KAPA) synthase

Shanghua Fan, De Feng Li, Da Cheng Wang, Joy Fleming, Hongtai Zhang, Ying Zhou, Lin Zhou, Jie Zhou, Tao Chen, Guanjun Chen, Xian En Zhang, Lijun Bi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The biotin biosynthesis pathway is an attractive target for development of novel drugs against mycobacterial pathogens, however there are as yet no suitable inhibitors that target this pathway in mycobacteria. 7-Keto-8-aminopelargonic acid synthase (KAPA synthase, BioF) is the enzyme which catalyzes the first committed step of the biotin synthesis pathway, but both its structure and function in mycobacteria remain unresolved. Here we present the crystal structure of Mycobacterium smegmatis BioF (MsBioF). The structure reveals an incomplete dimer, and the active site organization is similar to, but distinct from Escherichia coli 8-amino-7-oxononanoate synthase (EcAONS), the E. coli homologue of BioF. To investigate the influence of structural characteristics on the function of MsBioF, we deleted bioF in M. smegmatis and confirmed that BioF is required for growth in the absence of exogenous biotin. Based on structural and mutagenesis studies, we confirmed that pyridoxal 5′-phosphate (PLP) binding site residues His129, Lys235 and His200 are essential for MsBioF activity in vivo and residue Glu171 plays an important, but not essential role in MsBioF activity. The N-terminus (residues 1-37) is also essential for MsBioF activity in vivo. The structure and function of MsBioF reported here provides further insights for developing new anti-tuberculosis inhibitors aimed at the biotin synthesis pathway.

Original languageEnglish (US)
Pages (from-to)71-80
Number of pages10
JournalInternational Journal of Biochemistry and Cell Biology
Volume58
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

Fingerprint

8-amino-7-oxononanoate synthase
Mycobacterium smegmatis
Biotin
Escherichia coli
Mutagenesis
Pyridoxal Phosphate
Mycobacterium
Biosynthesis
Pathogens
Dimers
Crystal structure
Binding Sites
Enzymes
Catalytic Domain
Pharmaceutical Preparations
Tuberculosis

Keywords

  • 7-Keto-8-aminopelargonic acid (KAPA)synthase
  • Active site
  • Biotin synthesis pathway
  • Crystal structure
  • Mycobacterium smegmatis

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Structure and function of Mycobacterium smegmatis 7-keto-8-aminopelargonic acid (KAPA) synthase. / Fan, Shanghua; Li, De Feng; Wang, Da Cheng; Fleming, Joy; Zhang, Hongtai; Zhou, Ying; Zhou, Lin; Zhou, Jie; Chen, Tao; Chen, Guanjun; Zhang, Xian En; Bi, Lijun.

In: International Journal of Biochemistry and Cell Biology, Vol. 58, 01.01.2015, p. 71-80.

Research output: Contribution to journalArticle

Fan, S, Li, DF, Wang, DC, Fleming, J, Zhang, H, Zhou, Y, Zhou, L, Zhou, J, Chen, T, Chen, G, Zhang, XE & Bi, L 2015, 'Structure and function of Mycobacterium smegmatis 7-keto-8-aminopelargonic acid (KAPA) synthase', International Journal of Biochemistry and Cell Biology, vol. 58, pp. 71-80. https://doi.org/10.1016/j.biocel.2014.11.006
Fan, Shanghua ; Li, De Feng ; Wang, Da Cheng ; Fleming, Joy ; Zhang, Hongtai ; Zhou, Ying ; Zhou, Lin ; Zhou, Jie ; Chen, Tao ; Chen, Guanjun ; Zhang, Xian En ; Bi, Lijun. / Structure and function of Mycobacterium smegmatis 7-keto-8-aminopelargonic acid (KAPA) synthase. In: International Journal of Biochemistry and Cell Biology. 2015 ; Vol. 58. pp. 71-80.
@article{2b7468f8076f49f19b3b2c69b9f082ba,
title = "Structure and function of Mycobacterium smegmatis 7-keto-8-aminopelargonic acid (KAPA) synthase",
abstract = "The biotin biosynthesis pathway is an attractive target for development of novel drugs against mycobacterial pathogens, however there are as yet no suitable inhibitors that target this pathway in mycobacteria. 7-Keto-8-aminopelargonic acid synthase (KAPA synthase, BioF) is the enzyme which catalyzes the first committed step of the biotin synthesis pathway, but both its structure and function in mycobacteria remain unresolved. Here we present the crystal structure of Mycobacterium smegmatis BioF (MsBioF). The structure reveals an incomplete dimer, and the active site organization is similar to, but distinct from Escherichia coli 8-amino-7-oxononanoate synthase (EcAONS), the E. coli homologue of BioF. To investigate the influence of structural characteristics on the function of MsBioF, we deleted bioF in M. smegmatis and confirmed that BioF is required for growth in the absence of exogenous biotin. Based on structural and mutagenesis studies, we confirmed that pyridoxal 5′-phosphate (PLP) binding site residues His129, Lys235 and His200 are essential for MsBioF activity in vivo and residue Glu171 plays an important, but not essential role in MsBioF activity. The N-terminus (residues 1-37) is also essential for MsBioF activity in vivo. The structure and function of MsBioF reported here provides further insights for developing new anti-tuberculosis inhibitors aimed at the biotin synthesis pathway.",
keywords = "7-Keto-8-aminopelargonic acid (KAPA)synthase, Active site, Biotin synthesis pathway, Crystal structure, Mycobacterium smegmatis",
author = "Shanghua Fan and Li, {De Feng} and Wang, {Da Cheng} and Joy Fleming and Hongtai Zhang and Ying Zhou and Lin Zhou and Jie Zhou and Tao Chen and Guanjun Chen and Zhang, {Xian En} and Lijun Bi",
year = "2015",
month = "1",
day = "1",
doi = "10.1016/j.biocel.2014.11.006",
language = "English (US)",
volume = "58",
pages = "71--80",
journal = "International Journal of Biochemistry and Cell Biology",
issn = "1357-2725",
publisher = "Elsevier Limited",

}

TY - JOUR

T1 - Structure and function of Mycobacterium smegmatis 7-keto-8-aminopelargonic acid (KAPA) synthase

AU - Fan, Shanghua

AU - Li, De Feng

AU - Wang, Da Cheng

AU - Fleming, Joy

AU - Zhang, Hongtai

AU - Zhou, Ying

AU - Zhou, Lin

AU - Zhou, Jie

AU - Chen, Tao

AU - Chen, Guanjun

AU - Zhang, Xian En

AU - Bi, Lijun

PY - 2015/1/1

Y1 - 2015/1/1

N2 - The biotin biosynthesis pathway is an attractive target for development of novel drugs against mycobacterial pathogens, however there are as yet no suitable inhibitors that target this pathway in mycobacteria. 7-Keto-8-aminopelargonic acid synthase (KAPA synthase, BioF) is the enzyme which catalyzes the first committed step of the biotin synthesis pathway, but both its structure and function in mycobacteria remain unresolved. Here we present the crystal structure of Mycobacterium smegmatis BioF (MsBioF). The structure reveals an incomplete dimer, and the active site organization is similar to, but distinct from Escherichia coli 8-amino-7-oxononanoate synthase (EcAONS), the E. coli homologue of BioF. To investigate the influence of structural characteristics on the function of MsBioF, we deleted bioF in M. smegmatis and confirmed that BioF is required for growth in the absence of exogenous biotin. Based on structural and mutagenesis studies, we confirmed that pyridoxal 5′-phosphate (PLP) binding site residues His129, Lys235 and His200 are essential for MsBioF activity in vivo and residue Glu171 plays an important, but not essential role in MsBioF activity. The N-terminus (residues 1-37) is also essential for MsBioF activity in vivo. The structure and function of MsBioF reported here provides further insights for developing new anti-tuberculosis inhibitors aimed at the biotin synthesis pathway.

AB - The biotin biosynthesis pathway is an attractive target for development of novel drugs against mycobacterial pathogens, however there are as yet no suitable inhibitors that target this pathway in mycobacteria. 7-Keto-8-aminopelargonic acid synthase (KAPA synthase, BioF) is the enzyme which catalyzes the first committed step of the biotin synthesis pathway, but both its structure and function in mycobacteria remain unresolved. Here we present the crystal structure of Mycobacterium smegmatis BioF (MsBioF). The structure reveals an incomplete dimer, and the active site organization is similar to, but distinct from Escherichia coli 8-amino-7-oxononanoate synthase (EcAONS), the E. coli homologue of BioF. To investigate the influence of structural characteristics on the function of MsBioF, we deleted bioF in M. smegmatis and confirmed that BioF is required for growth in the absence of exogenous biotin. Based on structural and mutagenesis studies, we confirmed that pyridoxal 5′-phosphate (PLP) binding site residues His129, Lys235 and His200 are essential for MsBioF activity in vivo and residue Glu171 plays an important, but not essential role in MsBioF activity. The N-terminus (residues 1-37) is also essential for MsBioF activity in vivo. The structure and function of MsBioF reported here provides further insights for developing new anti-tuberculosis inhibitors aimed at the biotin synthesis pathway.

KW - 7-Keto-8-aminopelargonic acid (KAPA)synthase

KW - Active site

KW - Biotin synthesis pathway

KW - Crystal structure

KW - Mycobacterium smegmatis

UR - http://www.scopus.com/inward/record.url?scp=84912528184&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84912528184&partnerID=8YFLogxK

U2 - 10.1016/j.biocel.2014.11.006

DO - 10.1016/j.biocel.2014.11.006

M3 - Article

C2 - 25462832

AN - SCOPUS:84912528184

VL - 58

SP - 71

EP - 80

JO - International Journal of Biochemistry and Cell Biology

JF - International Journal of Biochemistry and Cell Biology

SN - 1357-2725

ER -