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Title: BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF THE 35-KDA PERIPLASMIC IRON-REGULATED PROTEIN FROM PASTEURELLA HAEMOLYTICA.

Author
item Belzer, Carol
item Tabatabai, Louisa
item Frank, Glynn

Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 11/10/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Pasteurella haemolytica A1 is a bacterial pathogen that causes bovine respiratory disease of cattle. Relatively little is known about the mechanism of iron acquisition of Pasteurella. A 35-kDa protein that may play a role in iron acquisition by P. haemolytica has been isolated, purified and characterized. P. haemolytica A1 was grown in RPMI medium without iron by established methods. The periplasmic proteins were extracted by using an osmotic shock procedure and also by using a hypertonic salt solution. The isolated proteins were purified as described previously. Molecular weight was determined by SDS-PAGE, matrix-assisted laser desorption (MALDI) mass spectrometry, HPLC-electrospray mass spectrometry (ES-MS), and analytical equilibrium velocity ultracentrifugation. The N-terminal sequence and amino acid composition were determined after blotting to polyvinylidene difluoride (PVDF) membrane. Secondary structure was determined by circular dichroism. The results show that the protein has a molecular weight of 35 kDa and that the native protein is a single polypeptide with a molecular weight of 35 kDa. The CD spectra indicate the protein contains primarily beta sheet structure.