Cloning, purification, crystallization and preliminary X-ray diffraction studies of Escherichia coli PapD-like protein (EcpD)

Nishant Kumar Pandey, Ravi Kant Pal, Kashyap Maruthi, Neel Sarovar Bhavesh

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Many Gram-negative bacteria are characterized by hair-like proteinaceous appendages on their surface known as fimbriae. In uropathogenic strains of Escherichia coli, fimbriae mediate attachment by binding to receptors on the host cell, often contributing to virulence and disease. E. coli PapD-like protein (EcpD) is a periplasmic chaperone that plays an important role in the proper folding and guiding of Yad fimbrial proteins to the outer membrane usher protein in a process known as pilus biogenesis. EcpD is essential for pilus biogenesis in uropathogenic E. coli and plays an important role in virulence. In the present study, EcpD was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 1.67 Å resolution and belonged to the orthorhombic space group C2221, with unit-cell parameters a = 100.3, b = 127.6, c = 45.9 Å. There was a single molecule in the asymmetric unit and the corresponding Matthews coefficient was calculated to be 3.02 Å 3 Da -1, with 59% solvent content. Initial phases were determined by molecular replacement.

Original languageEnglish (US)
Pages (from-to)954-957
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume68
Issue number8
DOIs
StatePublished - Aug 1 2012
Externally publishedYes

Fingerprint

Cloning
Escherichia
Crystallization
purification
X-Ray Diffraction
Uropathogenic Escherichia coli
Escherichia coli
Purification
Organism Cloning
crystallization
proteins
X ray diffraction
Virulence
virulence
biological evolution
diffraction
Proteins
x rays
Gram-Negative Bacteria
appendages

Keywords

  • chaperones
  • EcpD
  • PapD-like protein
  • pilus biogenesis
  • uropathogenic Escherichia coli

ASJC Scopus subject areas

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

Cite this

Cloning, purification, crystallization and preliminary X-ray diffraction studies of Escherichia coli PapD-like protein (EcpD). / Pandey, Nishant Kumar; Pal, Ravi Kant; Maruthi, Kashyap; Bhavesh, Neel Sarovar.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 68, No. 8, 01.08.2012, p. 954-957.

Research output: Contribution to journalArticle

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