Biosynthesis, processing and half-life of P-glycoprotein in a human multidrug-resistant KB cell

Akihiko Yoshimura, Yasuo Kuwazuru, Tomoyuki Sumizawa, Shun ichi Ikeda, Misako Ichikawa, Takashi Usagawa, Shin ichi Akiyama

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

The biosynthesis, processing, and half-life of the drug efflux pump, P-glycoprotein, were studied in human multidrug-resistant KB (KB-C2) cells selected for resistance to colchicine. An antibody directed against a synthetic oligopeptide corresponding to the amino-acid sequence (Glu-393-Lys-408) of P-glycoprotein from human mdrl cDNA was prepared in rabbits. With immunoblotting and immunoprecipitation, we detected a 140-170 kDa protein in KB-C2 cells but not in parental sensitive KB cells. KB-C2 cells made a 125 kDa precursor that was slowly processed (t 1 2 = 45 min) to the mature form of 140-150 kDa. The processing rate of P-glycoprotein was slower than that of low-density lipoprotein receptor. We detected another 160-180 kDa smear band, which might be a completely denatured form of P-glycoprotein. With immunoblotting, a minor band of high molecular mass (greater than 500 kDa) was also detected and this form increased after the cells were treated with chemical cross-linker, 1,5-difluoro-2,4-dinitrobenzene. The half-life of P-glycoprotein was long; no significant loss of P-glycoprotein was observed within 24 h after synthesis. Cells treated with tumicamycin produced a 120 kDa from of P-glycoprotein which was no longer processed but showed stability similar to that of the mature 140-150 kDa form. Agents that reverse multidrug resistance, phorbol ester and transport substrate did not affect the stability of P-glycoprotein.

Original languageEnglish
Pages (from-to)307-314
Number of pages8
JournalBBA - General Subjects
Volume992
Issue number3
DOIs
Publication statusPublished - 1989 Sep 15
Externally publishedYes

Fingerprint

KB Cells
Biosynthesis
P-Glycoprotein
Half-Life
Processing
Immunoblotting
Oligopeptides
LDL Receptors
Colchicine
Molecular mass
Multiple Drug Resistance
Phorbol Esters
Immunoprecipitation
Amino Acid Sequence
Complementary DNA
Pumps
Rabbits
Amino Acids
Antibodies
Substrates

Keywords

  • (Human)
  • (KB cell)
  • Multidrug resistance
  • P-glycoprotein

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Yoshimura, A., Kuwazuru, Y., Sumizawa, T., Ikeda, S. I., Ichikawa, M., Usagawa, T., & Akiyama, S. I. (1989). Biosynthesis, processing and half-life of P-glycoprotein in a human multidrug-resistant KB cell. BBA - General Subjects, 992(3), 307-314. https://doi.org/10.1016/0304-4165(89)90089-5

Biosynthesis, processing and half-life of P-glycoprotein in a human multidrug-resistant KB cell. / Yoshimura, Akihiko; Kuwazuru, Yasuo; Sumizawa, Tomoyuki; Ikeda, Shun ichi; Ichikawa, Misako; Usagawa, Takashi; Akiyama, Shin ichi.

In: BBA - General Subjects, Vol. 992, No. 3, 15.09.1989, p. 307-314.

Research output: Contribution to journalArticle

Yoshimura, A, Kuwazuru, Y, Sumizawa, T, Ikeda, SI, Ichikawa, M, Usagawa, T & Akiyama, SI 1989, 'Biosynthesis, processing and half-life of P-glycoprotein in a human multidrug-resistant KB cell', BBA - General Subjects, vol. 992, no. 3, pp. 307-314. https://doi.org/10.1016/0304-4165(89)90089-5
Yoshimura, Akihiko ; Kuwazuru, Yasuo ; Sumizawa, Tomoyuki ; Ikeda, Shun ichi ; Ichikawa, Misako ; Usagawa, Takashi ; Akiyama, Shin ichi. / Biosynthesis, processing and half-life of P-glycoprotein in a human multidrug-resistant KB cell. In: BBA - General Subjects. 1989 ; Vol. 992, No. 3. pp. 307-314.
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