The charge characterization of native and deglycosylated thyrotropin

Brooks Allen Keel, Robin L. Harms, Syed M. Amir

Research output: Contribution to journalArticle

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Abstract

Chromatofocusing was used to characterize the charge microbeterogeneity of crude pituitary, highly purified native bovine (b) and deglycosylated (dg) thyrotropin (TSH) preparations. Greater than 90% of crude pituitary TSH and native bTSH-I-1 bound to concanavalin-A (conA) columns while dgbTSH was excluded from the column. Extracts of ovine (o) pituitaries contained at least nine species (isohormones) of immunoreactive TSH when chromatofocused on pH 7.5-4 gradients. Highly purified native bTSH-I-1 displayed a similar elution profile. In contrast, dgbTSH eluted as a single homogeneous species with an apparent pI > 7.5. When subjected to chromatofocusing on a pH 10.5-7 gradient, 68% of crude pituitary oTSH and 96% of native bTSH-I-1 was bound to the column but could be eluted with NaCl indicating acidic species, while at least three peaks of dgbTSH could be resolved having apparent pI's of 9.12, 9.03 and 8.98. These data suggest that although removal of the carbohydrate moieties markedly alters the isohormone pattern of TSH, chemical deglycosylation does not completely eliminate the charge microbeterogeneity of bTSH.

Original languageEnglish (US)
Pages (from-to)151-163
Number of pages13
JournalEndocrine Research
Volume16
Issue number2
DOIs
StatePublished - Jan 1 1990
Externally publishedYes

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Concanavalin A
Sheep
Carbohydrates
deglycosylated thyrotropin

ASJC Scopus subject areas

  • Endocrinology

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The charge characterization of native and deglycosylated thyrotropin. / Keel, Brooks Allen; Harms, Robin L.; Amir, Syed M.

In: Endocrine Research, Vol. 16, No. 2, 01.01.1990, p. 151-163.

Research output: Contribution to journalArticle

Keel, Brooks Allen ; Harms, Robin L. ; Amir, Syed M. / The charge characterization of native and deglycosylated thyrotropin. In: Endocrine Research. 1990 ; Vol. 16, No. 2. pp. 151-163.
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