Induction of muscle protein degradation and weight loss by a tumor product.
Splenocytes from mice bearing a cachexia-inducing tumor (MAC16) have been fused with mouse myeloma cells to produce hybridomas, which have been cloned to produce antibody reactive to a material which copurified with a lipid-mobilizing factor isolated from the same tumor. The monoclonal antibody has been used to investigate factors potentially involved in the development of cachexia. The major protein detectable by immunoprecipitation of a partially purified lipid-mobilizing factor was M(r) 69,000, whereas Western blotting showed two bands of M(r) 69,000 and M(r) 24,000. Although the monoclonal antibody did not neutralize lipid-mobilizing activity in an in vitro assay, it did neutralize a serum factor capable of protein degradation in isolated gastrocnemius muscle. Affinity purification of MAC16 tumor homogenates using the monoclonal antibody yielded two immunoreactive bands of M(r) 69,000 and M(r) 24,000, which were further fractionated on a hydrophobic column (C8). This material was capable of inducing tyrosine release from isolated gastrocnemius muscle, and the effect could be blocked with the monoclonal antibody. The two immunoreactive bands from the hydrophobic column were capable of inducing weight loss in mice, whereas nonimmunoreactive fractions had no effect on body weight. The M(r) 24,000 species had a unique amino acid sequence, whereas the M(r) 69,000 species gave the same sequence as the M(r) 24,000 material, together with that for albumin. The M(r) 24,000 species contained carbohydrate, and lectin blotting showed a strong reaction with wheat germ and Erythrina crystagalli agglutinins. This suggests that the material is a glycoprotein or proteoglycan that shows strong binding affinity for albumin, possibly through the carbohydrate residues.
Todorov
P T
PT
Cancer Research Campaign Nutritional Biochemistry Research Group, Pharmaceutical Sciences Institute, Aston University, Birmingham, United Kingdom.
McDevitt
T M
TM
Cariuk
P
P
Coles
B
B
Deacon
M
M
Tisdale
M J
MJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000494:pharmacology; D015153:Blotting, Western; D002100:Cachexia / Q000209:etiology*; D002459:Cell Fusion; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008970:Molecular Weight; D009101:Multiple Myeloma / Q000276:immunology; D009124:Muscle Proteins / Q000187:drug effects* / Q000378:metabolism; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D009369:Neoplasms / Q000150:complications* / Q000378:metabolism; D013154:Spleen / Q000166:cytology
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D009124:Muscle Proteins; D009363:Neoplasm Proteins