ISOLATION AND STRUCTURE OF HUMAN CORTICOTROPIN (ACTH)<sup>1</sup>

Lee, Teh H.; Lerner, Aaron B.; Buettner-Janusch, Vina

doi:10.1021/ja01531a059

Cited by 38 publications

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“…2). The amino acid sequence of human y-trace compared to the sequences of human glucagon (24) and human corticotropin (25). Identical residues of the molecules are shown in boxes.…”

Section: Resultsmentioning

confidence: 99%

Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis.

Grubb¹,

Löfberg²

1982

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

287

127

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The amino acid sequence of human V-trace, a basic microprotein without known function, was determined by automated Edman degradations of the carboxynethylated polypeptide chain and of fragments obtained by cyanogen bromide treatment and tryptic digestion after blocking of lysine residues. The single polypeptide chain contained 120 residues, and the calculated Mr was 13,260. A proline residue at position 3 was partly hydroxylated. The presence of y-trace in a significant proportion of the cells in the anterior lobe of simian and human pituitary glands was demonstrated by immunohistochemical procedures with a rabbit antiserum against human V-trace. The tissue localization and amino acid sequence of V-trace indicated that this protein is connected with the peptidergic gastroenteropancreatic neuroendocrine system. Human y-trace is an alkaline, low Mr protein first described in 1961 as a constituent ofnormal cerebrospinal fluid and of urine from patients with renal failure (1-3). The biological role of ytrace is unknown, although the protein has been shown to occur in low concentration not only in cerebrospinal fluid and urine but also in plasma, saliva, and semen (4-6). A sensitive enzyme immunoassay for measurement of y-trace has been described (7). It allowed quantitation of the protein in all human fluids investigated and demonstrated that, in healthy adults, the concentration of y-trace in cerebrospinal fluid is 5.5 times that in plasma. Recent immunohistochemical studies of y-trace have shown it to be present in some human brain cortical neurons, in adrenal medulla, and in the A cells of the pancreatic islets (8-10). Some amino acid sequence similarities also have been found between human glucagon and the amino-terminal part of y-trace (8). Therefore, the question whether y-trace is related to the peptidergic gastroenteropancreatic neuroendocrine system has arisen. This report describes the complete amino acid sequence of y-trace and presents immunohistochemical evidence of the presence of the protein in a large number of cells in the pituitary gland. Experimental details of the work will be published separately.MATERIALS AND METHODS Isolation of V-Trace. Urine containing about 20 mg of y-trace per liter from patients with renal failure was collected in bottles containing the protease inhibitor benzamidinium chloride and the antimicrobial agent sodium azide. y-Trace was isolated from the pooled urine as described (8). Its purity was checked by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (11) and agarose gel electrophoresis (12).Structural Determination. Isolated y-trace was reduced and carboxymethylated with '4C-labeled reagent in 6 M guanidinium chloride. The carboxymethylated protein was then fragmented by cyanogen bromide treatment (13) or by tryptic digestion after reversible blocking ofamino groups by citraconylation (14). One ofthe cyanogen bromide fragments (no. 3) was further digested with Staphylococcus aureus V8 protease. On each occasion the fragments were separated by gel chro...

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“…2). The amino acid sequence of human y-trace compared to the sequences of human glucagon (24) and human corticotropin (25). Identical residues of the molecules are shown in boxes.…”

Section: Resultsmentioning

confidence: 99%

Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis.

Grubb¹,

Löfberg²

1982

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

287

127

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“…Porcine ACTH, synthetic human des-Ser ACTH-(2-39)-octatriakontapeptide and ACTH-(31-39)-nonapeptide of the sequence of Lee, Lerner & Buettner-Janusch (1961), were gifts from Mr P. Lloyd and Dr R. L. Colescott of Armour Pharma¬ ceutical Co., Eastbourne, England and Kankakee, Illinois, U.S.A. Collagenase was purchased from the Boerhringer Corp. (London) Ltd, and outdated human serum albumin was provided by the Lister Institute, Elstree, Herts. Na125I (catalogue number IMS-30) was obtained from the Radiochemical Centre, Amersham, Bucks.…”

Section: Methodsmentioning

confidence: 99%

Studies on the Binding of 125i-Labelled Corticotrophin to Isolated Rat Adrenocortical Cells

McIlhinney¹,

Schulster²

1975

Journal of Endocrinology

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Isolated adrenocortical cells, prepared by collagenase disaggregation of normal rat adrenal glands, have been used to study the binding characteristics of 125I-labelled corticotrophin (ACTH) of established biological activity. The binding of the labelled hormone to these whole cells was highly specific, only peptides possessing steroidogenic activity displaced the labelled hormone. Binding was rapid, being complete within 5 min of adding the hormone, and the amount of hormone bound remained constant for up to 20 min thereafter. Over the range 160-10000pg ACTH/ml, increased binding of the hormone was observed at all concentrations of hormone which stimulated steroidogenesis. However at levels of ACTH which stimulated maximal steroidogenesis there was no saturation of binding. This provides the first direct evidence for the existence of 'spare receptors' for ACTH on whole adrenocortical cells. Scatchard analysis of the binding data suggests that there are two types of receptor for ACTH in this preparation of cells. One receptor is of high affinity (dissociation constant equals 2-5 times 10-10 mol/l) with about 3000 sites/cell and the other is of lower affinity (dissociation constant equals 1 times 10-8 mol/l) with about 30000 sites/cell.

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“…It was later found in other species and found to be chemically identical to the porcine molecule (39,(126)(127)(128)(129). All known alp ha-MSH's are composed of the first 13 residues of ACTH; the N-terminal serine may be either acetylated or not acetylated (39).…”

Section: Alpha-mshmentioning

confidence: 94%

Chemistry and biosynthesis of pro-opiomelanocortin

Chrétien

Seidah

1981

Mol Cell Biochem

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Studies of lipotropins, melanotropins and endorphins on one hand, and of adrenocorticotropin on the other, has given rise to the concept of a multipotent precursor molecule recently renamed proopiomelanocortin. The preferential sites of cleavage of the precursor to produce its biologically active components are made of pairs of basic amino acid residues as described for the biosynthesis of beta-MSH and pro-insulin. Such structural feature is also found in other pro-hormone molecules. Pulse chase experiments and secretory studies carried out in both anterior and intermediate lobes of rat pituitary glands revealed the transformation of different forms of the precursor into different end-products, the anterior lobe producing preferentially ACTH and beta-LPH while the intermediate produces mainly the alpha-MSH and beta-endorphin. The multiple forms of precursors seem to differ in their carbohydrate content although at least two different gene products are still possible. The presence of similar peptides in the hypothalamus makes it highly probable that neuropeptides are biosynthesized with similar process. Thus the model of beta-LPH precursor, proposed as early as in 1967, is now applicable to the biosynthesis of all other neuropeptides. Major advances in this field are expected in the 1980s.

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ISOLATION AND STRUCTURE OF HUMAN CORTICOTROPIN (ACTH)¹

Cited by 38 publications

References 0 publications

Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis.

Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis.

Studies on the Binding of 125i-Labelled Corticotrophin to Isolated Rat Adrenocortical Cells

Chemistry and biosynthesis of pro-opiomelanocortin

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ISOLATION AND STRUCTURE OF HUMAN CORTICOTROPIN (ACTH)1

Cited by 38 publications

References 0 publications

Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis.

Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis.

Studies on the Binding of 125i-Labelled Corticotrophin to Isolated Rat Adrenocortical Cells

Chemistry and biosynthesis of pro-opiomelanocortin

Contact Info

Product

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ISOLATION AND STRUCTURE OF HUMAN CORTICOTROPIN (ACTH)¹