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Infection and Immunity, July 1999, p. 3181-3187, Vol. 67, No. 7
0019-9567/99/$04.00+0
Effects of Environmental pH on Membrane Proteins
in Borrelia burgdorferi
James A.
Carroll,1,*
Claude F.
Garon,1 and
Tom G.
Schwan2
Rocky Mountain Laboratories Microscopy
Branch1 and Laboratory of Microbial
Structure and Function,2 Rocky Mountain
Laboratories, National Institute of Allergy and Infectious
Diseases, Hamilton, Montana 59840
Received 5 January 1999/Returned for modification 18 February
1999/Accepted 6 April 1999
Borrelia burgdorferi, the causative agent of Lyme
disease, alternates between the microenvironments of the tick vector,
Ixodes scapularis, and a mammalian host. The environmental
conditions the spirochete encounters during its infectious cycle are
suspected to differ greatly in many aspects, including available
nutrients, temperature, and pH. Here we identify alterations in
the membrane protein profile, as determined by immunoblotting and
two-dimensional nonequilibrium pH gradient gel electrophoresis
(2D-NEPHGE), that occur in virulent B. burgdorferi B31 as the pH of the medium is altered. Initial
comparisons of cultures incubated at pHs 6.0, 7.0, and 8.0 yielded alterations in the expression of seven membrane proteins as
determined by probing with hyperimmune rabbit serum. Six of
these membrane proteins (54, 45, 44, 43, 35, and 24 kDa) were
either present in increased amounts in or solely expressed by
cultures incubated at pHs 6.0 and 7.0. The 24-kDa protein that decreased in expression at pH 8.0 was identified as outer surface protein C (OspC). In addition, a 42-kDa membrane protein increased in
amount in cultures incubated at pH 8.0. Similar changes were observed
with serum from a mouse infected by tick bite, with the recognition of
two additional bands (48 and 46 kDa) unique to pHs 6.0 and 7.0. When
membrane fractions were analyzed by 2D-NEPHGE, at least 37 changes in
the membrane protein profile between cells incubated at pHs 6.0, 7.0, and 8.0 were observed by immunoblotting and silver staining.
Environmental cues such as pH may prove important in the regulation of
virulence determinants and factors necessary for the
adaptation of B. burgdorferi to the tick or mammalian microcosm.
*
Corresponding author. Mailing address: 903 South 4th
St., Hamilton, MT 59840. Phone: (406) 363-9407. Fax: (406) 363-9371. E-mail: jcarroll{at}atlas.niaid.nih.gov.
Infection and Immunity, July 1999, p. 3181-3187, Vol. 67, No. 7
0019-9567/99/$04.00+0
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