Subcellular localization of pyruvate dehydrogenase dihydrolipoamide acetyltransferase in human intrahepatic biliary epithelial cells

Joplin, Ruth; Wallace, L L; Johnson, G. D.; Lindsay, J. Gordon; Yeaman, Stephen J.; Palmer, Jeremy M.; Strain, Alastair J.; Neuberger, J

doi:10.1002/path.1711760409

Cited by 34 publications

(8 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The distribution of rabbit anti-WMSYPDRTLRTS binding to sections of the liver was similar to the previously observed distribution of anti-PDC-E2 antibodies (11,12,24). Highest intensity of binding was associated with medium-sized interlobular ducts, the ducts primarily involved in PBC.…”

Section: Discussionsupporting

confidence: 86%

Random phage mimotopes recognized by monoclonal antibodies against the pyruvate dehydrogenase complex-E2 (PDC-E2).

Cha

Leung

Water

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Dihydrolipoamide acetyltransferase, the E2 component ofthe pyruvate dehydrogenase complex (PDC-E2), is the autoantigen most. commonly recognized by autoantibodies in primary biliary cirrhosis (PBC). We identified a peptide mimotope(s) of PDC-E2 by screening a phage-epitope library expressing random dodecapeptides in the pIll coat protein of fd phage using C355.1, a murine monoclonal antibody (mAb)that recognizes a conformation-dependent epitope in the inner lipoyl domain of PDC-E2 and uniquely stains the apical region of bile duct epithelium (BDE) only in patients with PBC. Eight different sequences were identified in 36 phage clones. WMSYPDRTLRTS was present in 29 clones; WESYPFRVGTSL, APKTYVSVSGMV, LTYVSL-QGRQGH, LDYVPLKHRHRH, AALWGVKVRHVS, KVLN-RIMAGVRH and GNVALVSSRVNA were singly represented. Three common amino acid motifs (W-SYP, TWS, and VRH) were shared among all peptide sequences. Competitive inhibition of the immunohistochemical staining of PBC BDE was performed by incubating the peptides WMSYPDRTLRTS, WESYPDRTLRTS, APKTWSVSGMV, and AALWGVKV-RHVS with either C355.1 or a second PDC-E2-specific mAb, C150.1. Both mAbs were originally generated to PDC-E2 but map to distinct regions of PDC-E2. Two of the peptides, although selected by reaction with C355.1, strongly inhibited the staining of BDE by C150.1, whereas the peptide APK-TYVSVSGMV consistently inhibited the staining of C355.1 on biliary duct epithelium more strongly than the typical mitochondrial staining of hepatocytes. Rabbit sera raised against the peptide WMSYPDRTLRTS stained BDE of livers and isolated bile duct epithelial cells of PBC patients more intensively than controls. The rabbit sera stained all size ducts in normals, but only small/medium-sized ductules in PBC livers.These studies provide evidence that the antigen present in BDE is a molecular mimic of PDC-E2, and not PDC-E2 itself.

show abstract

Section: Discussionsupporting

confidence: 86%

Random phage mimotopes recognized by monoclonal antibodies against the pyruvate dehydrogenase complex-E2 (PDC-E2).

Cha

Leung

Water

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…However, one of the eight antibodies (C355.1) showed greater intensity and specificity for the apical region of biliary epithelial cells only from patients with PBC. Similar results have also been obtained with affinity-purified anti-PDC-E2 and combinatorial (13)(14)(15). Attempts to characterize this antigen have failed for a variety of reasons, including the small number and low viability of biliary epithelial cells in diseased livers.…”

supporting

confidence: 66%

Cryptic Antigenic Determinants on the Extracellular Pyruvate Dehydrogenase Complex/Mimeotope Found in Primary Biliary Cirrhosis

Yip

Water

Gershwin

et al. 1996

Journal of Biological Chemistry

View full text Add to dashboard Cite

Affinity mass spectrometry (AMS) was used to evaluate the structural diversity of the E2 component of pyruvate dehydrogenase complex (PDC) in normal and diseased liver cells, including those from patients with the autoimmune disease primary biliary cirrhosis (PBC). Two different antibodies to PDC-E2, the immunodominant mitochondrial autoantigen in patients with PBC, were used. AMS was performed directly on frozen liver sections and purified bile duct epithelial cells. Mass spectrometric signals associated with the molecular recognition of PBC-specific antigenic determinants were enhanced by an in situ enzyme-linked signal amplification process. Samples from patients with PBC gave strong positive signals for the antigen(s) recognized by the monoclonal antibody C355.1. Conversely, tissues from normal and disease controls showed only a minimal signal. AMS was used to identify specific antigenic determinants within the E2 component of PDC for comparison with unknown antigenic determinants observed by affinity capture with C355.1 monoclonal antibody from PBC samples. PDC components bound to C355.1 were mapped and identified by mass before dissociation from the E2 component. A similar approach was used to identify unknown antigenic determinants associated with PBC. We believe AMS may be an important new approach with wide application to the identification of molecules associated with a number of disease states.

show abstract

“…A subset of mitochondrial proteins was identified in the biotinylation experiment. A number of these proteins, including ATPase 73, pyruvate dehydrogenase 74 and the voltage‐dependent anion channel 1 28, have been localized to the plasma membrane surface in various models and their identification here suggests that they occupy a similar location in O. viverrini . An ultrastructural investigation showed that the O. viverrini tegument is rich in mitochondria that, in many cases, were proximate to the cytoplasmic side of the tegumental plasma membrane 72.…”

Section: Resultsmentioning

confidence: 77%

The secreted and surface proteomes of the adult stage of the carcinogenic human liver fluke Opisthorchis viverrini

et al. 2010

View full text Add to dashboard Cite

Infection with the human liver fluke, Opisthorchis viverrini, is a serious public health problem in Thailand, Laos and nearby locations in Southeast Asia. Both experimental and epidemiological evidence strongly implicate liver fluke infection in the etiology of one of the liver cancer subtypes, cholangiocarcinoma (CCA). To identify parasite proteins critical for liver fluke survival and the etiology of CCA, OFFGEL electrophoresis and Multiple reaction monitoring were employed to characterize 300 parasite proteins from the O. viverrini excretory/secretory products (ES) and, utilizing selective labeling and sequential solubilization, from the host-exposed tegument. The ES included a complex mixture of proteins that have been associated with cancers, including proteases of different mechanistic classes and orthologues of mammalian growth factors and anti-apoptotic proteins. Also identified was a cysteine protease inhibitor which, in other helminth pathogens, induces nitric oxide production by macrophages, and, hence may contribute to malignant transformation of inflamed cells. Proteases in ES included cathepsins, calpain and a protein with homology to autoimmune prostatitis antigen 2. More than 160 tegumental proteins were identified using sequential solubilization of isolated teguments, and a subset of these was localized to the surface membrane of the tegument by labeling living flukes with biotin and confirming surface localization with fluorescence microscopy. These included annexins, which are potential immuno-modulators, and orthologues of the schistosomiasis vaccine antigens Sm29 and tetraspanin-2. Novel roles in pathogenesis were suggested for the tegument-host interface since more than ten biotinylated surface proteins had no homologues in the public databases. The O. viverrini proteins identified here provide an extensive catalogue of novel leads for research on the pathogenesis of opisthorchiasis and the development of novel interventions for this disease and CCA, as well as providing a scaffold for sequencing the genome of this fluke.

show abstract

Subcellular localization of pyruvate dehydrogenase dihydrolipoamide acetyltransferase in human intrahepatic biliary epithelial cells

Cited by 34 publications

References 24 publications

Random phage mimotopes recognized by monoclonal antibodies against the pyruvate dehydrogenase complex-E2 (PDC-E2).

Random phage mimotopes recognized by monoclonal antibodies against the pyruvate dehydrogenase complex-E2 (PDC-E2).

Cryptic Antigenic Determinants on the Extracellular Pyruvate Dehydrogenase Complex/Mimeotope Found in Primary Biliary Cirrhosis

The secreted and surface proteomes of the adult stage of the carcinogenic human liver fluke Opisthorchis viverrini

Contact Info

Product

Resources

About