Human Parathyroid Hormone: Immunological Properties of the Amino Terminus and of Synthetic Fragments

1974 ◽  
Vol 47 (6) ◽  
pp. 567-576 ◽  
Author(s):  
G. N. Hendy ◽  
P. M. Barling ◽  
J. L. H. O'Riordan
Keyword(s):  
Parathyroid Hormone ◽  
Synthetic Peptides ◽  
Terminal Region ◽  
Amino Terminus ◽  
Human Parathyroid Hormone ◽  
Immunological Properties ◽  
Immunological Assay ◽  
Amino Terminal

1. Immunological assay systems specific to the amino-terminal region of parathyroid hormone were used to study the properties of human parathyroid hormone (HPTH 1-84) and of synthetic peptides. One of these peptides (HPTH 1-34A) had the sequence proposed by Niall, Sauer, Jacobs, Keutmann, Segre, O'Riordan, Aurbach & Potts (1974). The others (HPTH 1-34B and C, synthesized by different methods) had the sequence proposed by Brewer, Fairwell, Ronan, Sizemore & Arnaud (1972). 2. HPTH 1-84 and HPTH 1-34A behaved identically in a number of these systems. In some systems, especially those using antisera obtained by immunizing with HPTH 1-34A, differences were, however, found. These may be due to conformational or other physical differences or to the presence of an immunologically incomplete site in the fragment. 3. In systems in which HPTH 1–84 and HPTH 1-34A behaved identically the peptides with the sequence proposed by Brewer et al. (1972) were of low reactivity by 100–2000-fold.

DEVELOPMENT OF HOMOLOGOUS IMMUNOLOGICAL ASSAYS FOR HUMAN PARATHYROID HORMONE

1980 ◽  
Vol 85 (1) ◽  
pp. 161-170 ◽  
Author(s):  
R. M. MANNING ◽  
G. N. HENDY ◽  
S. E. PAPAPOULOS ◽  
J. L. H. O'RIORDAN
Keyword(s):  
Parathyroid Hormone ◽  
Trichloroacetic Acid ◽  
Terminal Region ◽  
Human Parathyroid Hormone ◽  
Antibody Assay ◽  
Amino Terminal ◽  
High Affinity ◽  
Carboxy Terminal Region ◽  
Immunological Assays ◽  
Carboxy Terminal

SUMMARY Antisera to a trichloroacetic-acid precipitate of human parathyroid hormone (PTH) were produced in goats. Two of these antisera (G36 and G31) were of high affinity, and the bovine and porcine hormones were less reactive. Synthetic peptides containing the amino-terminal region of human PTH reacted with both antisera; the 1–34 peptide (PTH-(1–34)), with the sequence proposed by Niall, Sauer, Jacobs, Keutmann, Segre, O'Riordan, Aurbach & Potts in 1974, was more reactive than that having the sequence proposed by Brewer, Fairwell, Ronan, Sizemore & Arnaud in 1972. The antisera were further characterized with a number of other native and synthetic fragments of human PTH and reacted poorly with fragments from the carboxy-terminal region of the molecule. Since the amino-terminal fragments did not account for all the immunoreactivity, it is assumed that the antisera had some recognition sites for the central part of the molecule. Highly purified human PTH-(1–84) was labelled with 125I and radioimmunoassays were developed using this tracer and antiserum G36. To avoid the problems associated with labelling human PTH with 125I, a labelled antibody assay was developed with G36 and an immunoadsorbent consisting of human PTH-(1–34) (sequence of Niall et al.) coupled to cellulose. A sensitive homologous amino-terminal specific assay was developed in this way.

IMMUNOLOGICAL PROPERTIES OF SYNTHETIC HUMAN PARATHYROID HORMONE 53–84 FRAGMENT

1979 ◽  
Vol 80 (1) ◽  
pp. 153-155 ◽  
Author(s):  
G. N. HENDY ◽  
R. M. MANNING ◽  
M. ROSENBLATT ◽  
G. W. TREGEAR ◽  
H. T. KEUTMANN ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Synthetic Peptide ◽  
Terminal Region ◽  
Human Parathyroid Hormone ◽  
Enzymic Digestion ◽  
Immunological Properties ◽  
Carboxyl Terminal

The immunological properties of a synthetic peptide comprising the carboxyl-terminal 53–84 region of human parathyroid hormone (PTH) have been studied. The immunoreactivity of the synthetic human PTH-(53–84) peptide paralleled that of a 53–84 fragment of the native human hormone prepared by enzymic digestion, in both a standard radioimmunoassay, which was not region-specific, and also a radioimmunoassay specific for the carboxyl-terminal region of PTH. However, in both types of radioimmunoassay the synthetic human PTH-(53–84) peptide was four to five times more reactive than the native human PTH-(53–84) fragment.

Amino-Terminal Modifications of Human Parathyroid Hormone (PTH) Selectively Alter Phospholipase C Signaling via the Type 1 PTH Receptor:  Implications for Design of Signal-Specific PTH Ligands†

Biochemistry ◽  
1999 ◽  
Vol 38 (41) ◽  
pp. 13453-13460 ◽  
Author(s):  
Hisashi Takasu ◽  
Thomas J. Gardella ◽  
Michael D. Luck ◽  
John T. Potts ◽  
F. Richard Bringhurst
Keyword(s):  
Parathyroid Hormone ◽  
Phospholipase C ◽  
Human Parathyroid Hormone ◽  
Amino Terminal

ACTH and adrenal aerobic glycolysis. I: Effects of O-nitrophenylsulphenyl and other ACTH analogues, vasoactive intestinal peptide and human parathyroid hormone(1–34) on lactic acid, steroid and cyclic AMP production by mouse adrenocortical cells

1987 ◽  
Vol 115 (1) ◽  
pp. 61-69 ◽  
Author(s):  
J. Hinson ◽  
M. K. Birmingham
Keyword(s):  
Parathyroid Hormone ◽  
Cyclic Amp ◽  
Vasoactive Intestinal Peptide ◽  
Aerobic Glycolysis ◽  
Fold Increase ◽  
Terminal Segment ◽  
Human Parathyroid Hormone ◽  
Experimental Conditions ◽  
Adrenal Cells ◽  
Amino Terminal

ABSTRACT The structural requirements in the ACTH molecule for evocation of the glycolytic response in suspensions of mouse adrenal cells were investigated by examining the effects of analogues containing modifications at positions 8,9 and 10 and of peptides containing homologies with the amino-terminal segment of ACTH. Introduction of a nitrophenylsulphenyl (NPS) group into the tryptophan moiety at position 9 of ACTH(1–24) greatly reduced both the potency and the capacity for maximal glycolytic response. It also virtually abolished cyclic AMP formation. In contrast, the capacity for a maximal steroidogenic response remained unimpaired in the NPS derivative, although steroidogenic potency was reduced to 0·4% of that of ACTH(1–24). Replacement of the tryptophan moiety with phenylalanine had intermediate inhibitory effects on glycolysis and steroid output; replacement with alanine virtually abolished both these responses. Replacement of arginine in position 8 with lysine in the Phe9 analogue caused a fifty-fold increase in glycolytic potency, but rendered it steroidogenically inactive. Cyclic AMP production was abolished in the Ala9 analogue and greatly impaired in the Phe9 and Lys8,Phe9 analogues. Replacement of the glycine moiety in position 10 with l-alanine, d-alanine, β-alanine or α-aminoisobutyric acid had little or no effect on steroidogenic or glycolytic capacity, although potency was reduced with all substitutions excepting l-alanine. Vasoactive intestinal peptide, which contains homologies with positions 3, 7, 15 and 16 of ACTH, proved completely inactive in dispersed mouse adrenal cells under our experimental conditions, in contrast to human parathyroid hormone(1–34) (hPTH (1–34)) which contains homologies with positions 3, 4, 5, 9 and 11 and was steroidogenic at the lowest concentration tested (0·1 nmol/l), eliciting an eleven-fold increase in steroid production, a response which might be physiologically relevant. It induced near maximal steroidogenesis at a concentration of 10 nmol/l, without affecting cyclic AMP production, and stimulated glycolysis at concentrations above 10 nmol/l, accompanied by a slight rise in cyclic AMP levels. The examples of dissociation between glycolysis and steroidogenesis suggest that different receptors may mediate the two responses. The examples of increased steroidogenesis unaccompanied by a rise in cyclic AMP accord with the concept that cyclic AMP is not an obligatory second messenger for the steroidogenic response. J. Endocr. (1987) 115, 61–69

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