Identification of carbohydrate deficient transferrin forms by MALDI-TOF mass spectrometry and lectin ELISA

Peter, Jochen; Unverzagt, Carlo; Engel, Wolf-Dieter; Renauer, Doris; Seidel, Christoph; Hösel, Wolfgang

doi:10.1016/s0304-4165(97)00135-9

Cited by 52 publications

(24 citation statements)

References 19 publications

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“…Mass Number Analysis by Matrix-assisted Laser Desorption and Ionization (MALDI) Spectrometry-Mass number analyses were carried out according to the method of Peter et al (29) by using a Kompact MALDI 4 (Shimadzu Corp.) operated in linear time of flight mode. Ionization was accomplished by nitrogen laser at a wavelength of 337 nm for 3 ns.…”

Section: Methodsmentioning

confidence: 99%

Prophenoloxidase-activating Enzyme of the Silkworm, Bombyx mori

Satoh

Horii

Ochiai

et al. 1999

Journal of Biological Chemistry

214

143

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Prophenoloxidase-activating enzyme (PPAE) was purified to homogeneity as judged by SDS-polyacrylamide gel electrophoresis from larval cuticles of the silkworm, Bombyx mori. The purified PPAE preparation was shown to be a mixture of the isozymes of PPAE (PPAE-I and PPAE-II), which were eluted at different retention times in reversed-phase high performance liquid chromatography. PPAE-I and PPAE-II seemed to be post translationally modified isozymes and/or allelic variants. Both PPAE isozymes were proteins composed of two polypeptides (heavy and light chains) that are linked by disulfide linkage(s) and glycosylated serine proteases. The results of cDNA cloning, peptide mapping, and amino acid sequencing of PPAE revealed that PPAE is synthesized as prepro-PPAE with 441 amino acid residues and is activated from pro-PPAE by cleavage of a peptide bond between Lys 152 and Ile 153 . The homology search showed 36.9% identity of PPAE to easter, which is a serine protease involved in dorsoventral pattern formation in the Drosophila embryo, and indicated the presence of two consecutive clip-like domains in the light chain. A single copy of the PPAE gene was suggested to be present in the silkworm genome. In the fifth instar larvae, PPAE transcripts were detected in the integument, hemocytes, and salivary glands but not in the fat body or mid gut. A polypeptide cross-reactive to mono-specific anti-PPAE/IgG was transiently detected in the extract of eggs between 1 and 3 h after they were laid.

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Section: Methodsmentioning

confidence: 99%

Prophenoloxidase-activating Enzyme of the Silkworm, Bombyx mori

Satoh

Horii

Ochiai

et al. 1999

Journal of Biological Chemistry

214

143

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“…In the ensuing decades, CDT became the most heavily validated biomarker in the alcohol field (Salaspuro, 1999;Bortolotti et al, 2006). Further refinements have identified a specific isoform, disialotransferrin with a single N-glycan chain (Landberg et al, 1995;Peter et al, 1998;Flahaut et al, 2003), as the more specific marker for chronic heavy alcohol drinking. This disialo-isoform can be quantified by high-performance liquid chromatography (HPLC) (Helander et al, 2003(Helander et al, , 2010 and expressed as a percentage of total transferrin, which is referred to in this manuscript as %dCDT.…”

Section: Introductionmentioning

confidence: 99%

Relationship of Abnormal Chromatographic Pattern for Carbohydrate-Deficient Transferrin with Severe Liver Disease

Stewart

Reuben

Anton

2016

Alcohol and Alcoholism

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Aims: Serum carbohydrate-deficient transferrin (CDT) is a validated test for chronic heavy alcohol drinking, but CDT abnormalities have been associated with liver disease, limiting its use in these patients. We report here on the association between poor chromatographic resolution of disialotransferrin from trisialotransferrin (the so-called 'di-tri bridging') and liver disease severity and etiology. Methods: Subjects were patients in whom detailed clinical data, including histology results, were available on their existing liver diseases (n=139). Percent disialo-CDT (%dCDT) was measured by high-performance liquid chromatography, and the risks for di-tri bridging associated with cirrhosis, with and without adjustment for alcohol use and alcohol-related liver disease, were estimated. Results: Di-tri bridging was present in 22/73 (30%) cirrhotic subjects and 7/66 (11%) non-cirrhotic subjects. The unadjusted risk for di-tri bridging in cirrhotics relative to non-cirrhotics was 3.6 (95% confidence interval 1.4-9.2). Adjustment for alcohol-related liver disease and current drinking had little effect on this estimate (adjusted odds ratio 3.4), and neither alcohol-related liver disease nor current drinking were independently associated with di-tri bridging after accounting for the effect of cirrhosis. Conclusions: The presence of di-tri bridging was associated with cirrhosis in individuals with both alcohol-related and non-alcoholic liver disease, although most cirrhotic subjects did not exhibit di-tri bridging. When di-tri bridging is seen in individuals being tested for chronic heavy drinking, investigation for cirrhosis should be considered. Short summary: There are known liver-disease-associated abnormalities in CDT. In this study, we found that such abnormalities were strongly associated with cirrhosis rather than less-advanced disease, but were only clinically evident in 30% of cirrhotics. Abnormalities also occurred in severe hepatitis without cirrhosis and were not specific for liver disease etiology.

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“…This important point needs further investigation. However, there is evidence (obtained by two independent analytical methods) of the complete lack of both N-glycans in at least some of the asialotransferrin molecules that are present in serum after chronic alcohol abuse (2,10,11 ). In this case, the analyte asialotransferrin is clearly defined and does not allow individual interpretation.…”

Section: To the Editormentioning

confidence: 99%

“…With a complete lack of both transferrin N-glycans (2,10,11 ), the structure of asialotransferrin is different from the structure(s) of the non-CDT isoforms. Thus, there is the possibility of raising a specific antibody that binds strongly to the analyte (asialotransferrin), e.g., to an epitope near or at the original point of attachment of the carbohydrate chain(s).…”

Section: To the Editormentioning

confidence: 99%

Asialotransferrin—An Alternative to Carbohydrate-deficient Transferrin?

Arndt¹

2003

Clinical Chemistry

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Identification of carbohydrate deficient transferrin forms by MALDI-TOF mass spectrometry and lectin ELISA

Cited by 52 publications

References 19 publications

Prophenoloxidase-activating Enzyme of the Silkworm, Bombyx mori

Prophenoloxidase-activating Enzyme of the Silkworm, Bombyx mori

Relationship of Abnormal Chromatographic Pattern for Carbohydrate-Deficient Transferrin with Severe Liver Disease

Asialotransferrin—An Alternative to Carbohydrate-deficient Transferrin?

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