Lateral heterogeneity in the cytoplasmic membrane of Bacillus subtilis was found by using density gradient centrifugation. Crude membranes (CM) present in the whole cell lysate were separated into three fractions of increasing density (F, CI, CII). Substantial difference exists in the amount of protein recovered from these fractions, the relative ratio being 15 : 35 : 50. The qualitative protein composition (by SDS-PAGE) of the fractions varies markedly as well. The lipid components extracted from the fractions are also distributed in different proportions, viz. 40 : 40 : 20. The spectrum of fatty acids (FA), detected in lipids of F fraction and analyzed by GC-MS exhibits the same profile as that found in CM; in contrast, fractions CI and CII undergo extensive FA reconstruction. Thermotropic behavior of fractions measured by the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene indicates significant variations of microviscosity (r(s)) within the F, CI and CII fractions. The protein-to-lipid ratio plays evidently a key role in affecting the physical state of the cytoplasmic membrane. Microdomains of different density coexist in the membrane and exhibit heterogeneity in both chemical composition and "physical state"; the increased de novo synthesis of FA induced by the cold exclusively in fractions CI and CII indicates correlation with an altered physiological state of bacterial metabolism.

Department of Genetics and Microbiology Faculty of Science Charles University Prague Prague

Grant č.156/2006

Lit.: 28

000      

03422naa 2200421 a 4500

001      

bmc07503494

003      

CZ-PrNML

005      

20111210121740.0

008      

080507s2007 xr e eng||

009      

AR

040    __

$a ABA008 $b cze $c ABA008 $d ABA008 $e AACR2

041    0_

$a eng

044    __

$a xr

100    1_

$a Toman, O.

245    10

$a Detection of lateral heterogeneity in the cytoplasmic membrane of Bacillus subtilis / $c O. Toman, F. Le Hégarat, Jaroslava Svobodová

314    __

$a Department of Genetics and Microbiology, Faculty of Science, Charles University in Prague, Prague

500    __

$a Grant č.156/2006

504    __

$a Lit.: 28

520    9_

$a Lateral heterogeneity in the cytoplasmic membrane of Bacillus subtilis was found by using density gradient centrifugation. Crude membranes (CM) present in the whole cell lysate were separated into three fractions of increasing density (F, CI, CII). Substantial difference exists in the amount of protein recovered from these fractions, the relative ratio being 15 : 35 : 50. The qualitative protein composition (by SDS-PAGE) of the fractions varies markedly as well. The lipid components extracted from the fractions are also distributed in different proportions, viz. 40 : 40 : 20. The spectrum of fatty acids (FA), detected in lipids of F fraction and analyzed by GC-MS exhibits the same profile as that found in CM; in contrast, fractions CI and CII undergo extensive FA reconstruction. Thermotropic behavior of fractions measured by the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene indicates significant variations of microviscosity (r(s)) within the F, CI and CII fractions. The protein-to-lipid ratio plays evidently a key role in affecting the physical state of the cytoplasmic membrane. Microdomains of different density coexist in the membrane and exhibit heterogeneity in both chemical composition and "physical state"; the increased de novo synthesis of FA induced by the cold exclusively in fractions CI and CII indicates correlation with an altered physiological state of bacterial metabolism.

650    _2

$a Bacillus subtilis $x izolace a purifikace $x růst a vývoj $7 D001412

650    _2

$a cytoplazmatické struktury $x chemie $x mikrobiologie $7 D022083

650    _2

$a membránové mikrodomény $x chemie $x mikrobiologie $7 D021962

650    _2

$a membránové proteiny $x chemie $x izolace a purifikace $7 D008565

650    _2

$a membránové lipidy $x chemie $x izolace a purifikace $7 D008563

650    _2

$a fluorescenční polarizace $x metody $x využití $7 D005454

650    _2

$a elektroforéza v polyakrylamidovém gelu $x metody $x využití $7 D004591

650    _2

$a chromatografie plynová $x metody $x využití $7 D002849

650    _2

$a finanční podpora výzkumu jako téma $7 D012109

700    1_

$a Le Hégarat, F.

700    1_

$a Svobodová, Jaroslava

773    0_

$w MED00011005 $t Folia microbiologica $g Roč. 52, č. 4 (2007), s. 339-345 $x 0015-5632

910    __

$a ABA008 $b A 2403 $c 561 $y 1

990    __

$a 20080507092633 $b ABA008

991    __

$a 20080513084409 $b ABA008

999    __

$a ok $b bmc $g 619115 $s 471548

BAS    __

$a 3

BMC    __

$a 2007 $b 52 $c 4 $d 339-345 $i 0015-5632 $m Folia microbiologica $x MED00011005

LZP    __

$a 2008-3/mkal