The Lipid Nature of a Tumour‐Associated Autoantigen from a Chemically Induced Rat Hepatoma

Lando, P.; Berzins, Klavs; Perlmann, Peter

doi:10.1111/j.1365-3083.1982.tb00637.x

Cited by 5 publications

(3 citation statements)

References 15 publications

(11 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The increase in phenanthroline-detectable copper in serum or synovial fluid stored at 4°C is accompanied by a decrease in ferroxidase I activity, suggesting that the copper originates from degradation of caeruloplasmin. The immunological activity of caeruloplasmin does not decrease, however, perhaps because the degraded protein is still immunoreactive (see, e.g., Lando et al, 1982). Release of Cu(II) ions may accelerate lipid peroxidation in the fluids (Gutteridge, 1977;Halliwell & Gutteridge, 1984), which would account for the parallel increases in diene conjugation and-thiobarbituric acid-reactive material.…”

Section: Discussionmentioning

confidence: 99%

The behaviour of caeruloplasmin in stored human extracellular fluids in relation to ferroxidase II activity, lipid peroxidation and phenanthroline-detectable copper

Gutteridge¹,

Winyard²,

Blake³

et al. 1985

Biochemical Journal

View full text Add to dashboard Cite

No Cu(II) ion is measurable in human serum or synovial fluid by the phenanthroline assay. On storage of human serum or synovial fluid at 4 degrees C, phenanthroline-detectable copper appears, lipid peroxidation occurs, ferroxidase I activity declines and ferroxidase II activity rises, yet there is no fall in immunologically detectable caeruloplasmin. Storage of body fluids at -20 degrees C or -70 degrees C slows, but does not prevent, these deteriorative changes. It is suggested that the presence of low-molecular-mass Cu(II) ion complexes, ferroxidase II activity, "cytotoxic factors' and "immunosuppressive factors' in body fluids may be, in part or in whole, an artifact of the storage and handling of the fluids. A report [Blake, Blann, Bacon, Farr, Gutteridge & Halliwell (1983) Clin. Sci. 64, 551-553] that the caeruloplasmin present in rheumatoid synovial fluid is deficient in ferroxidase activity is shown to be such an artifact. It is strongly recommended that all such experiments be performed upon freshly taken fluid samples.

show abstract

Section: Discussionmentioning

confidence: 99%

The behaviour of caeruloplasmin in stored human extracellular fluids in relation to ferroxidase II activity, lipid peroxidation and phenanthroline-detectable copper

Gutteridge¹,

Winyard²,

Blake³

et al. 1985

Biochemical Journal

View full text Add to dashboard Cite

show abstract

“…The dialyzed and centrifuged semisynthetic Mb (70 nmol) was applied to a column (20 x 1.2 cm) of Sepharose 4B (Pharmacia) to which 250 nmol of goat anti-Mb antibody (for preparation, see ref. 15) was covalently attached. The column was, washed at 10 ml/hr at 40C until the optical absorbance of the eluate indicated no further passage of protein as monitored at 280 nm.…”

Section: Methodsmentioning

confidence: 99%

Semisynthesis of sperm whale myoglobin by fragment condensation.

Simmerman¹,

Wang²,

Horwitz³

et al. 1982

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

View full text Add to dashboard Cite

Reconstruction of the sperm whale myoglobin structure was accomplished by a series of aqueous condensations of suitably protected synthetic myoglobin fragments to a large fragment prepared from the native protein. Reaction of NANT9-acetimidomyoglobin with 3-bromo-2-(2-nitrophenylsulfenyl)skatole (BNPS-skatole) yielded the fragment corresponding to residues 15-153. The covalent structure was reformed by sequential coupling of the N-hydroxysuccinimide esters of o-nitrophenylsulfenyl-L-tryptophan (residue 14) and selectively protected peptides corresponding to residues 1-5 and 6-13, which were synthesized by the solid-phase method and removed from the resin by methoxide-catalyzed methanolysis. A mixed aqueous solvent system containing methanol and N,N,N',N'-tetrakis(2-hydroxypropyl)ethylenediamine/trifluoroacetic acid buffer (Quadrol) solubilized the peptide and protein fragments during the condensations. Replacement of the heme moiety and immunoaffinity chromatography made possible the isolation and purification of the reconstructed native molecule. The development of this nondestructive synthetic procedure allows investigation of the structural and functional significance of individual residues by isotopic enrichment or selective amino acid substitutions.Studies using selective, consecutive removal and substitution ofthe NH2-terminal residues ofMb by specific degradation and resynthesis (1, 2) have indicated the subtle contribution of the NH2-terminal charge and the uncharged side chains ofresidues 1 and 2 to the conformation and stability of the native molecule (3). To examine further the effects of synthetic sequence alteration with respect to stability, conformation, and electrostatic and hydrophobic interactions, a method for creating internal sequence variations must be developed. A promising approach is that of semisynthesis, a technique whereby the natural product is selectively cleaved and a fragment is isolated that is suitable for rebuilding the native structure or an analog.Previous attempts at partial synthesis with Mb have utilized tryptic or CNBr fragments in which citraconyl or maleyl protecting groups were used on the e-NH2 groups (4, 5). The current strategy exploits the oxidative lability of the tryptophans at positions 7 and 14 ofthe Mb sequence by the use of3-bromo-2-(nitrophenylsulfenyl)skatole (BNPS-skatole; skatole = 3-methylindole) to effect cleavage, yielding the fragment corresponding to residues 15-153 (designated fragment 15-153) in which the E-NH2 groups have been protected previously by reaction with methyl acetimidate (1). Sequential coupling of the N-hydroxysuccinimide (HOSu) esters of o-nitrophenylsulfenyl-L-tryptophan (NPS-Trp; residue 14) and the selectively protected peptides corresponding to residues 1-5 and 6-13 (designated peptides 1-5 and 6-13) regenerated the native sequence. Isolation of reconstructed Mb capable of correctly positioning the heme moiety was accomplished by passage through successive affinity columns on which the native semisynthetic protein was se...

show abstract

“…Serum antibodies have most often been evaluated on the basis of binding to established cell lines or antigen preparations. Tumour specificity and the presence of antigen in normal tissue have been elucidated by absorption tests with various normal and tumour tissues [3,21] or by direct binding to selected nontumour cells. On the basis of results from this kind of experiment it has been stated that responses against individually unique tumour antigens are rare and that the major part of the serum antibody response is directed against antigens shared among many tumours and also present on nonmalignant cells (normal tissue antigens) [27].…”

Section: Introductionmentioning

confidence: 99%

Rat serum antibodies binding to high-molecular-weight glycoprotein(s) on syngeneic colon carcinomas, demonstrated by competition with rat monoclonal antibody

Jansson

Brodin

Sjögren

1989

Cancer Immunol Immunother

View full text Add to dashboard Cite

Sera from rats immunized to syngeneic 1,2-dimethylhydrazine colon carcinomas were analyzed for their ability to inhibit the binding of a syngeneic rat IgM monoclonal antibody (10B12) specific for high-molecular-weight glycoprotein(s) from rat colon carcinoma. Immunization with irradiated tumour cells or with tumour tissue extracts resulted in the appearance of a strong inhibiting activity. Sera of animals with established growing tumours and of females shortly after partus also inhibited binding of the monoclonal antibody, while unimmunized animals or animals immunized with irrelevant antigens had no inhibiting antibodies in their sera. Dimethylhydrazine-treated animals showed an increased titer of antibodies binding to the high-molecular-weight glycoprotein, but showed no inhibition of binding of the 10B12 monoclonal antibody. The syngeneic 10B12 rat antibody obviously does not reflect a rare event captured from a hyperimmune animal by the hybridoma technique but rather represents an antibody specificity frequently appearing in the immune response to tumours expressing the high-molecular-weight glycoprotein.

show abstract

The Lipid Nature of a Tumour‐Associated Autoantigen from a Chemically Induced Rat Hepatoma

Cited by 5 publications

References 15 publications

The behaviour of caeruloplasmin in stored human extracellular fluids in relation to ferroxidase II activity, lipid peroxidation and phenanthroline-detectable copper

The behaviour of caeruloplasmin in stored human extracellular fluids in relation to ferroxidase II activity, lipid peroxidation and phenanthroline-detectable copper

Semisynthesis of sperm whale myoglobin by fragment condensation.

Rat serum antibodies binding to high-molecular-weight glycoprotein(s) on syngeneic colon carcinomas, demonstrated by competition with rat monoclonal antibody

Contact Info

Product

Resources

About