Hemorphin Peptides are Released from Hemoglobin by Cathepsin D. Radioimmunoassay Against the C-part of V-V-hemorphin-7: An Alternative Assay for the Cathepsin D Activity

Garreau, Isabelle; Cucumel, K.; Dagouassat, Nathalie; Zhao, Qing; Cupo, A.; Piot, Jean–Marie

doi:10.1016/s0196-9781(96)00284-7

Cited by 23 publications

(16 citation statements)

References 32 publications

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“…18). It is also involved in the conversion of prohormones and precursor forms of biologically active peptides to active forms [132] and contributes to their inactivation. (Table 3).…”

Section: Role In Protein Degradationmentioning

confidence: 99%

See 1 more Smart Citation

Human cathepsin D.

Minarowska

Gacko²,

Karwowska³

et al. 2008

Folia Histochem Cytobiol.

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“…18). It is also involved in the conversion of prohormones and precursor forms of biologically active peptides to active forms [132] and contributes to their inactivation. (Table 3).…”

Section: Role In Protein Degradationmentioning

confidence: 99%

“…(Table 3). From hemoglobin, cathepsin D releases biologically active hemorphins [131,132]. Thus, it plays a regulatory role.…”

Section: Role In Protein Degradationmentioning

confidence: 99%

Human cathepsin D.

Minarowska

Gacko²,

Karwowska³

et al. 2008

Folia Histochem Cytobiol.

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“…Cohen et al measured serum levels of LVV-H7 from breast cancer patients by an ELISA technique [1,7]. As antibodies directed towards the C-terminal part of the peptide sequence were of limited quality in terms of selectivity [1], no stability studies or metabolite profiling had been performed using immunological methods. LVV-H7 and truncated derivatives lacking the first and/or the last amino acid were detected in adrenal and pheochromocytoma tumor tissue by MALDI-TOF MS and quantified in micromolar concentrations by LC-MS [4].…”

Section: Introductionmentioning

confidence: 99%

Comparative in vitro degradation of the human hemorphin LVV‐H7 in mammalian plasma analysed by capillary zone electrophoresis and mass spectrometry

John¹,

Schulz²,

Forssmann

2007

Biopharm & Drug Disp

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The human hemorphin LVV-H7 (L32VVYPWTQRF41) is a hemoglobin-beta, -gamma, -delta or -epsilon chain derived cationic decapeptide of the micro-opioid receptor binding family. It exhibits potential pharmacological value relevant, for example, for blood pressure regulation, learning performance and Alzheimer's disease. The regulatory potency is strictly dependent on the length of the amino acid sequence which is sensitive towards proteinases from tissues and plasma. To analyse LVV-H7 in vitro degradation in mammalian plasma, a novel multi-component quantitative capillary zone electrophoretic (CZE) procedure was applied, combined with qualitative metabolite profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In all types of plasma, LVV-H7 was N-terminally truncated generating four metabolites (M1-M4) with an intact C-terminus: M1 (V33VYPWTQRF41), M2 (V34YPWTQRF41), M3 (Y35PWTQRF41) and M4 (W37TQRF41). In EDTA plasma these degradation products were detected exclusively, whereas in citrate and heparin plasma four further metabolites appeared resulting from additional C-terminal cleavage of the dipeptide R40F41: M5 (L32VVYPWTQ39), M6 (V33VYPWTQ39), M7 (V34YPWTQ39) and M8 (Y35PWTQ39). In the presence of selective proteinase inhibitors aminopeptidase M and angiotensin-converting enzyme (for N- and C-terminal truncation, respectively) were identified as plasma enzymes responsible for hemorphin degradation. Furthermore, striking inter-mammalian species distinctions were detected revealing strongly differing degradation velocities but similar metabolite patterns.

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“…Bovine hemoglobin was prepared from erythrocytes as described previously [17]. Lysosomes (200 μg of protein) from fraction F 3 were incubated with hemoglobin (0.2% w/v) in a final volume of 500 μl of 4 mM sodium citrate buffer, pH 4.5 at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…Standard, samples, tracer and antiserum were diluted in RIA buffer. Radioimmunoassay procedure for VV‐hemorphin‐7 was based on the precipitation with polyethylene glycol (PEG) and was conducted as described in a previous paper [17]. Briefly, the fractions from the RP‐HPLC stage were lyophilized and reconstituted in 1 ml of RIA buffer.…”

Section: Methodsmentioning

confidence: 99%

Proteolytic degradation of hemoglobin by endogenous lysosomal proteases gives rise to bioactive peptides: hemorphins

et al. 1999

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Hemorphin generation by mice peritoneal macrophages has been recently reported, nevertheless no conclusive data exist to localize clearly the macrophage proteolytic activity implicated in their generation. Because lysosomes are believed to be the main site of degradation in the endocytic pathway, we have studied their potential implication in the generation of hemorphins from hemoglobin. When this protein is submitted to purified rat liver lysosomes, an early generation of hemorphin-7-related peptides, detected by a radioimmunoassay, was observed. These peptides seemed to be relatively stable during the first hours of hydrolysis.z 1999 Federation of European Biochemical Societies.

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Hemorphin Peptides are Released from Hemoglobin by Cathepsin D. Radioimmunoassay Against the C-part of V-V-hemorphin-7: An Alternative Assay for the Cathepsin D Activity

Cited by 23 publications

References 32 publications

Human cathepsin D.

Human cathepsin D.

Comparative in vitro degradation of the human hemorphin LVV‐H7 in mammalian plasma analysed by capillary zone electrophoresis and mass spectrometry

Proteolytic degradation of hemoglobin by endogenous lysosomal proteases gives rise to bioactive peptides: hemorphins

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