scholarly journals Analysis of phagosomal proteomes: From latex-bead to bacterial phagosomes

PROTEOMICS ◽  
2010 ◽  
Vol 10 (22) ◽  
pp. 4098-4116 ◽  
Author(s):  
Qingbo Li ◽  
Chinnaswamy Jagannath ◽  
Prahlad K. Rao ◽  
Christopher R. Singh ◽  
Giovanni Lostumbo
Keyword(s):  
Latex Bead

scholarly journals Phosphoinositides Regulate Membrane-dependent Actin Assembly by Latex Bead Phagosomes

2002 ◽  
Vol 13 (4) ◽  
pp. 1190-1202 ◽  
Author(s):  
Hélène Defacque ◽  
Evelyne Bos ◽  
Boyan Garvalov ◽  
Cécile Barret ◽  
Christian Roy ◽  
...  
Keyword(s):  
Binding Sites ◽  
Kinase Activity ◽  
Kinase Inhibitors ◽  
Membrane Surface ◽  
Latex Bead ◽  
Actin Assembly ◽  
Wild Type ◽  
Membrane Surfaces ◽  
Organelle Membrane

Actin assembly on membrane surfaces is an elusive process in which several phosphoinositides (PIPs) have been implicated. We have reconstituted actin assembly using a defined membrane surface, the latex bead phagosome (LBP), and shown that the PI(4,5)P2-binding proteins ezrin and/or moesin were essential for this process ( Defacque et al., 2000b ). Here, we provide several lines of evidence that both preexisting and newly synthesized PI(4,5)P2, and probably PI(4)P, are essential for phagosomal actin assembly; only these PIPs were routinely synthesized from ATP during in vitro actin assembly. Treatment of LBP with phospholipase C or with adenosine, an inhibitor of type II PI 4-kinase, as well as preincubation with anti-PI(4)P or anti-PI(4,5)P2 antibodies all inhibited this process. Incorporation of extra PI(4)P or PI(4,5)P2 into the LBP membrane led to a fivefold increase in the number of phagosomes that assemble actin. An ezrin mutant mutated in the PI(4,5)P2-binding sites was less efficient in binding to LBPs and in reconstituting actin assembly than wild-type ezrin. Our data show that PI 4- and PI 5-kinase, and under some conditions also PI 3-kinase, activities are present on LBPs and can be activated by ATP, even in the absence of GTP or cytosolic components. However, PI 3-kinase activity is not required for actin assembly, because the process was not affected by PI 3-kinase inhibitors. We suggest that the ezrin-dependent actin assembly on the LBP membrane may require active turnover of D4 and D5 PIPs on the organelle membrane.

Isolation of Latex Bead-and Mycobacteria-Containing Phagosomes

Cell Biology ◽  
2006 ◽  
pp. 57-62
Author(s):  
M KUHNEL ◽  
E ANES ◽  
G GRIFFITHS
Keyword(s):  
Latex Bead

scholarly journals New Latex Bead Agglutination Assay for Differential Diagnosis of Cattle Infected with Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis

2004 ◽  
Vol 11 (6) ◽  
pp. 1070-1074 ◽  
Author(s):  
Hye Cheong Koo ◽  
Yong Ho Park ◽  
Jongsam Ahn ◽  
W. Ray Waters ◽  
Mary Jo Hamilton ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
Mycobacterium Bovis ◽  
Mycobacterium Avium ◽  
Present Report ◽  
Area Under The Curve ◽  
High Specificity ◽  
Culture Method ◽  
Latex Bead ◽  
Characteristic Analysis ◽  
Agglutination Assay

ABSTRACT Extensive studies have shown that the current assays used to identify cattle infected with Mycobacterium bovis or Mycobacterium avium subsp. paratuberculosis are not sufficiently sensitive and specific to detect all infected animals, especially animals recently infected with the pathogens. In the present report we show that these limitations might be overcome with a latex bead agglutination assay (LBAA). With the specific immunodominant epitope (ESAT6-p) of M. bovis, we developed an LBAA and enzyme immunoassay (EIA) for that purpose and compared them with the “gold standard” culture method and skin test for their efficacy in detecting bovine tuberculosis. When sera from control healthy cows (n = 10), M. avium subsp. paratuberculosis-positive cattle (naturally infected, n = 16; experimentally infected, n = 8), and M. bovis-positive cattle (naturally infected, n = 49;experimentally infected, n = 20) were applied to an EIA and an LBAA developed with ESAT6-p, the two tests showed similar sensitivity (97.1% by EIA, 95.7% by LBAA), high specificity (94.2% by EIA, 100% by LBAA), and a positive correlation (kappa value, 0.85; correlation rate, 93.2%; correlation coefficient, 0.64). Receiver operating characteristic analysis of EIA results and comparison with the culture method determined a suitable cutoff value at 0.469, with an area under the curve of 0.991 (95% confidence interval, 0.977 to 1.0). As LBAA didn't show any positive reactions with sera from uninfected control cows or M. avium subsp. paratuberculosis-infected cattle, which were confirmed to be free of M. bovis by culture or PCR, LBAA using the ESAT6-p can be a rapid and useful M. bovis diagnostic assay. The data suggest that rapid, sensitive, and specific assays can be developed with peptides containing immunodominant epitopes present in proteins uniquely expressed in M. bovis or M. avium subsp. paratuberculosis for differential diagnosis of cattle infected with M. bovis or M. avium subsp. paratuberculosis.

Latex bead immobilisation in PDMS matrix for the detection of p53 gene point mutation and anti-HIV-1 capsid protein antibodies

2004 ◽  
Vol 381 (5) ◽  
pp. 1019-1024 ◽  
Author(s):  
Christophe A. Marquette ◽  
Agn�s Degiuli ◽  
Emmanuelle Imbert-Laurenceau ◽  
Francois Mallet ◽  
Carole Chaix ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Point Mutation ◽  
P53 Gene ◽  
Latex Bead ◽  
Anti Hiv ◽  
Hiv 1
Sign in / Sign up

Export Citation Format

Share Document