Tyrosine 67 in the epidermal growth factor-like domain of tissue-type plasminogen activator is important for clearance by a specific hepatic receptor

Rhonda Bassel-Duby, Nan Yan Jiang, Thomas Bittick, Edwin Madison, Dianna McGookey, Kim Orth, Ralph Shohet, Joseph Sambrook, Mary Jane Gething

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Abstract

Human tissue-type plasminogen activator (t-PA) is cleared rapidly from the circulation by hepatic receptors, one of which recognizes a site in the epidermal growth factor-like domain of the molecule. To define this site more precisely, we have used oligonucleotide-mediated mutagenesis to introduce amino acid substitutions at specific positions located in turns that connect antiparallel β-sheets in the epidermal growth factor-like domain. Mutated t-PA proteins with amino acid substitutions of the tyrosine residue at position 67 showed markedly lower rates of endocytosis and degradation by cultured cells of the rat hepatoma (H4) line that express a specific receptor for t-PA, and their half-life in the circulation of rats was extended significantly because of a reduction in the rate of the rapid α-phase of clearance. The enzymatic properties and fibrinolytic activity of these mutants in vitro were not significantly different from those of wild-type t-PA. We conclude that tyrosine 67 comprises a key determinant in the clearance of t-PA by a specific hepatic receptor.

Original languageEnglish (US)
Pages (from-to)9668-9677
Number of pages10
JournalJournal of Biological Chemistry
Volume267
Issue number14
StatePublished - May 15 1992

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Plasminogen Activators
Tissue Plasminogen Activator
Epidermal Growth Factor
Tyrosine
Liver
Amino Acid Substitution
Rats
Substitution reactions
Liver Circulation
Amino Acids
Mutagenesis
Endocytosis
Oligonucleotides
Half-Life
Hepatocellular Carcinoma
Cultured Cells
Cells
Degradation
Molecules
Proteins

ASJC Scopus subject areas

  • Biochemistry

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Tyrosine 67 in the epidermal growth factor-like domain of tissue-type plasminogen activator is important for clearance by a specific hepatic receptor. / Bassel-Duby, Rhonda; Jiang, Nan Yan; Bittick, Thomas; Madison, Edwin; McGookey, Dianna; Orth, Kim; Shohet, Ralph; Sambrook, Joseph; Gething, Mary Jane.

In: Journal of Biological Chemistry, Vol. 267, No. 14, 15.05.1992, p. 9668-9677.

Research output: Contribution to journalArticle

Bassel-Duby, Rhonda ; Jiang, Nan Yan ; Bittick, Thomas ; Madison, Edwin ; McGookey, Dianna ; Orth, Kim ; Shohet, Ralph ; Sambrook, Joseph ; Gething, Mary Jane. / Tyrosine 67 in the epidermal growth factor-like domain of tissue-type plasminogen activator is important for clearance by a specific hepatic receptor. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 14. pp. 9668-9677.
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AU - Jiang, Nan Yan

AU - Bittick, Thomas

AU - Madison, Edwin

AU - McGookey, Dianna

AU - Orth, Kim

AU - Shohet, Ralph

AU - Sambrook, Joseph

AU - Gething, Mary Jane

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