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Growth-associated modifications of low-molecular-weight thiols and protein sulfhydryls in human bronchial fibroblasts.
J Cell Physiol. 1990 Apr; 143(1):165-71.JC

Abstract

The thiol redox status of cultured human bronchial fibroblasts has been characterized at various growth conditions using thiol-reactive monobromobimane, with or without the combination of dithiotreitol, a strong reducing agent. This procedure has enabled measurement of the cellular content of reduced glutathione (GSH), total glutathione equivalents, cysteine, total cysteine equivalents, protein sulfhydryls, protein disulfides, and mixed disulfides. Passage of cells with trypsin perturbs the cellular thiol homeostasis and causes a 50% decrease in the GSH content, whereas the total cysteine content is subsequently increased severalfold during cell attachment. During subsequent culture, transient severalfold increased levels of GSH, protein-bound thiols, and protein disulfides are reached, whereas the total cysteine content gradually declines. These changes in the redox balance of both low-molecular-weight thiols and protein-bound thiols correlate with cell proliferation and mostly precede the major growth phase. When the onset of proliferation is inhibited by maintenance of cells in medium containing decreased amounts of serum, the GSH content remains significantly increased. Subsequent stimulation of growth by addition of serum results in decreased GSH levels at the onset of proliferation. In thiol-depleted medium, proliferation is also inhibited, whereas GSH levels are increased to a lesser extent than in complete medium. Exposure to buthionine sulfoximine inhibits growth, prevents GSH synthesis, and results in accumulation of total cysteine, protein-bound cysteine, and protein disulfides. For extracellular cystine, variable rates of cellular uptake correlate with the initial increase in the total cysteine content observed following subculture and with the GSH peak that precedes active proliferation. The results strongly suggest that specific fluctuations in the cellular redox balance of both free low-molecular-weight thiols and protein sulfhydryls are involved in growth regulation of normal human fibroblasts.

Authors+Show Affiliations

Department of Toxicology, Karolinska Institutet, Stockholm, Sweden.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

2318904

Citation

Atzori, L, et al. "Growth-associated Modifications of Low-molecular-weight Thiols and Protein Sulfhydryls in Human Bronchial Fibroblasts." Journal of Cellular Physiology, vol. 143, no. 1, 1990, pp. 165-71.
Atzori L, Dypbukt JM, Sundqvist K, et al. Growth-associated modifications of low-molecular-weight thiols and protein sulfhydryls in human bronchial fibroblasts. J Cell Physiol. 1990;143(1):165-71.
Atzori, L., Dypbukt, J. M., Sundqvist, K., Cotgreave, I., Edman, C. C., Moldéus, P., & Grafström, R. C. (1990). Growth-associated modifications of low-molecular-weight thiols and protein sulfhydryls in human bronchial fibroblasts. Journal of Cellular Physiology, 143(1), 165-71.
Atzori L, et al. Growth-associated Modifications of Low-molecular-weight Thiols and Protein Sulfhydryls in Human Bronchial Fibroblasts. J Cell Physiol. 1990;143(1):165-71. PubMed PMID: 2318904.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Growth-associated modifications of low-molecular-weight thiols and protein sulfhydryls in human bronchial fibroblasts. AU - Atzori,L, AU - Dypbukt,J M, AU - Sundqvist,K, AU - Cotgreave,I, AU - Edman,C C, AU - Moldéus,P, AU - Grafström,R C, PY - 1990/4/1/pubmed PY - 1990/4/1/medline PY - 1990/4/1/entrez SP - 165 EP - 71 JF - Journal of cellular physiology JO - J Cell Physiol VL - 143 IS - 1 N2 - The thiol redox status of cultured human bronchial fibroblasts has been characterized at various growth conditions using thiol-reactive monobromobimane, with or without the combination of dithiotreitol, a strong reducing agent. This procedure has enabled measurement of the cellular content of reduced glutathione (GSH), total glutathione equivalents, cysteine, total cysteine equivalents, protein sulfhydryls, protein disulfides, and mixed disulfides. Passage of cells with trypsin perturbs the cellular thiol homeostasis and causes a 50% decrease in the GSH content, whereas the total cysteine content is subsequently increased severalfold during cell attachment. During subsequent culture, transient severalfold increased levels of GSH, protein-bound thiols, and protein disulfides are reached, whereas the total cysteine content gradually declines. These changes in the redox balance of both low-molecular-weight thiols and protein-bound thiols correlate with cell proliferation and mostly precede the major growth phase. When the onset of proliferation is inhibited by maintenance of cells in medium containing decreased amounts of serum, the GSH content remains significantly increased. Subsequent stimulation of growth by addition of serum results in decreased GSH levels at the onset of proliferation. In thiol-depleted medium, proliferation is also inhibited, whereas GSH levels are increased to a lesser extent than in complete medium. Exposure to buthionine sulfoximine inhibits growth, prevents GSH synthesis, and results in accumulation of total cysteine, protein-bound cysteine, and protein disulfides. For extracellular cystine, variable rates of cellular uptake correlate with the initial increase in the total cysteine content observed following subculture and with the GSH peak that precedes active proliferation. The results strongly suggest that specific fluctuations in the cellular redox balance of both free low-molecular-weight thiols and protein sulfhydryls are involved in growth regulation of normal human fibroblasts. SN - 0021-9541 UR - https://www.unboundmedicine.com/medline/citation/2318904/Growth_associated_modifications_of_low_molecular_weight_thiols_and_protein_sulfhydryls_in_human_bronchial_fibroblasts_ L2 - https://doi.org/10.1002/jcp.1041430123 DB - PRIME DP - Unbound Medicine ER -