Characterization of a bioactive 15 kDa fragment produced by proteolytic cleavage of chicken growth hormone.
There is evidence for a cleaved form of GH in the chicken pituitary gland. A 25 kDa band of immunoreactive-(ir-)GH, as well as the 22 kDa monomeric form and some oligomeric forms were observed when purified GH or fresh pituitary extract were subjected to SDS-PAGE under nonreducing conditions. Under reducing conditions, the 25 kDa ir-GH was no longer observed, being replaced by a 15 kDa band, consistent with reduction of the disulfide bridges of the cleaved form. The type of protease involved was investigated using exogenous proteases and monomeric cGH. Cleaved forms of chicken GH were generated by thrombin or collagenase. The site of cleavage was found in position Arg133-Gly134 as revealed by sequencing the fragments produced. The NH2-terminal sequence of 40 amino acid residues in the 15 kDa form was identical to that of the rcGH and analysis of the remaining 7 kDa fragment showed an exact identity with positions 134-140 of cGH structure. The thrombin cleaved GH and the 15 kDa form showed reduced activity (0.8% and 0.5% of GH, respectively) in a radioreceptor assay employing a chicken liver membrane preparation. However, this fragment had a clear bioactivity in an angiogenic bioassay and was capable to inhibit the activity of deiodinase type III in the chicken liver.
Arámburo
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Centro de Neurobiologia, Universidad Nacional Autónoma de México, Querétaro. aramburo@servidor.unam.mx
Carranza
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Reyes
M
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Luna
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Martinez-Coria
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H
Berúmen
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L
Scanes
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CG
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002645:Chickens / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism; D019281:Dimerization; D004220:Disulfides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D013006:Growth Hormone / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D007453:Iodide Peroxidase / Q000037:antagonists & inhibitors; D008099:Liver / Q000648:ultrastructure; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D010902:Pituitary Gland / Q000737:chemistry; D011869:Radioligand Assay; D013917:Thrombin / Q000378:metabolism
D004220:Disulfides; D010446:Peptide Fragments; D013006:Growth Hormone; D007453:Iodide Peroxidase; D013917:Thrombin; D017364:Collagenases