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 language | English (US) |
|---|---|
| Pages (from-to) | 954-957 |
| Number of pages | 4 |
| Journal | Acta Crystallographica Section F: Structural Biology and Crystallization Communications |
| Volume | 68 |
| Issue number | 8 |
| DOIs | |
| State | Published - Aug 1 2012 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Cloning, purification, crystallization and preliminary X-ray diffraction studies of Escherichia coli PapD-like protein (EcpD)
AU - Pandey, Nishant Kumar
AU - Pal, Ravi Kant
AU - Maruthi, Kashyap
AU - Bhavesh, Neel Sarovar
PY - 2012/8/1
Y1 - 2012/8/1
N2 - 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.
AB - 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.
KW - chaperones
KW - EcpD
KW - PapD-like protein
KW - pilus biogenesis
KW - uropathogenic Escherichia coli
UR - http://www.scopus.com/inward/record.url?scp=84864869866&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84864869866&partnerID=8YFLogxK
U2 - 10.1107/S1744309112027364
DO - 10.1107/S1744309112027364
M3 - Article
C2 - 22869131
AN - SCOPUS:84864869866
VL - 68
SP - 954
EP - 957
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
SN - 1744-3091
IS - 8
ER -