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J A Biggins

Publications and source records attributed to J A Biggins.

27 records · Page 2Linked to original sources
PubMed1982

Corticotrophin-related peptides in the intermediate lobe of the rodent pituitary gland: characterization by high performance liquid chromatography and radioimmunoassay.

High performance liquid chromatography (HPLC) and radioimmunoassay have been used to characterize corticotrophin-related peptides in extracts of the intermediate lobe of the rat and mouse pituitary gland. Multiple peaks have been observed, which resemble corticotrophin-like intermediate lobe peptide (CLIP) in that they cross-react with antisera raised against the COOH-terminal region of corticotrophin (ACTH) but not against NH2-terminal directed antisera. These CLIP-like peptides were released from the incubated neurointermediate lobe and their secretion was inhibited in the presence of dopamine. Heterogeneity of peptide species was also observed with antisera raised against alpha-MSH. Multiple peaks of CLIP and alpha-MSH-like material were identified in pituitary extracts from the mouse and levels were elevated in the genetically obese (ob/ob) animal. The nature and possible functions of multiple forms of intermediate lobe peptides are discussed.

Adrenocorticotropic Hormone↗
PubMed1982

Mechanism of luteinizing hormone-releasing hormone degradation by subcellular fractions of rat hypothalamus and pituitary.

The pathway of LH-RH degradation by two subcellular fractions (a soluble fraction and a 25 000 X g particulate fraction) of rat hypothalamus, pituitary and cerebral cortex has been studied using high performance liquid chromatography and amino acid analysis to identify the breakdown products. The primary cleavage point in the Tyr5-Gly6 bond giving [1-5] LH-RH and [6-10] LH-RH. In the presence of dithiothreitol, cleavage of LH-RH also occurred at the Pro9-Gly10 bond giving [1-9] LH-RH. The fragment [1-5] LH-RH is further degraded sequentially from the C-terminus and [1-4] LH-RH, [1-3] LH-RH, tyrosine and tryptophan were identified. The other major fragment, [6-10] LH-RH, is rapidly broken down, the only intermediate product positively identified being Arg-Pro.

Animals↗
PubMed1981

Characterization and determination of neuropeptides by high-performance liquid chromatography and radioimmunoassay.

A method is described for the separation and analysis of a variety of neuropeptides using reversed-phase high-performance liquid chromatography coupled with radioimmunoassay. The solvent system (an acetonitrile gradient containing 0.08% trifluoroacetic acid) allows UV detection at 206 nm, gives good resolution and, by being volatile, is readily compatible with radioimmunoassay. Three applications of the method are described: (a) thyrotropin releasing hormone immunoreactivity in the rat brain has been characterized; (b) ACTH immunoreactivity in the rat pituitary pars intermedia has been resolved into its component peptides; (c) degradation of luteinizing hormone releasing hormone in vitro has been followed.

Adrenocorticotropic Hormone↗
PubMed1981

Degradation of luteinizing hormone-releasing hormone by serum and plasma in vitro.

Luteinizing hormone-releasing hormone (LH-RH) is degraded in vitro by serum and plasma from several species (human, rat, guinea-pig and cattle). Separation of the degradation products by high-performance liquid chromatography (HPLC) followed by amino acid analysis and radioimmunoassay showed that the main sites of cleavage are the Trp3-Ser4 and Tyr5-Gly6 bonds. Two peptidases are responsible since the cleavage at Trp3-Ser4 can be selectively inhibited by EDTA. In human plasma, the peptidase responsible for Trp3-Ser4 hydrolysis has a Km of 2.9 . 10(-4) M and V of 30 nmol/h per ml plasma. The half-life in vitro of LH-RH in serum and plasma from various species ranges from 3 h (guinea-pig) to 9.8 h (human). The peptidase cleaving LH-RH at Tyr5-Gly6 is present as an impurity in some commercial bovine serum and plasma albumins. Such contamination may have important practical implications for work involving peptide assays where albumins are used as carrier proteins.

Amino Acids↗
PubMed1978

Circulating antibody to myelin basic protein in relapsing-remitting multiple sclerosis? A comparative group and sequential study by radioimmunoassay.

Sera from multiple sclerosis patients with relapsing-remitting disease and normal subjects were tested for antibody to myelin basic protein by a sensitive radioimmunoassay. The results showed a marginally decreased titre in multiple sclerosis superimposed on a seasonal variation. There was no correlation with the clinical state of the patients. Results are discussed briefly in relation to humoral antibody function in multiple sclerosis and experimental autoimmune encephalitis.

Adult↗
PubMedDate not supplied

Failure of gel filtration and HPLC purification to increase levels of immunoreactive TRH in rat brain extracts.

It has recently been reported [4] that purification of acidified methanol extracts of rat cerebellum by gel filtration and high performance liquid chromatography (HPLC) caused a fifty-fold increase in immunoreactive thyrotrophin-releasing hormone (ir-TRH). However either using Sephadex LH20 or SepPak separation prior to HPLC we have failed to find a significant increase in ir-TRH in rat cerebellar or hypothalamic extracts. We conclude that it is valid to measure TRH by radioimmunoassay in unpurified methanolic extracts of brain tissue.

Animals↗
PubMedDate not supplied

Removal of Arg1 and Phe22 from CLIP (ACTH18-39) by rodent pituitary and blood peptidases.

The major product on incubation of CLIP (ACTH18-39) with rat and mouse serum, rat plasma and whole blood, and soluble extracts of rat pituitary is [des-Arg1]-CLIP (ACTH19-39) while [des-Phe22]-CLIP (ACTH18-38) is the major product with pituitary particulate fraction. In both cases, p-chloromercuribenzoate-sensitive, metal-dependent peptidase activity appears to be responsible for the cleavage. The serum enzyme may be related to proline aminopeptidase. Material coeluting with [des-Arg]-CLIP on two HPLC solvent gradients is present in the superfusion media from neurointermediate lobes of genetically obese (ob/ob) mice but is not present in acid extracts of the lobe. This suggests that postsecretory processing of CLIP may involve removal of the N-terminal Arg residue.

Adrenocorticotropic Hormone↗