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Identification of human milk α-lactalbumin as a cell growth inhibitor

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Summary

A growth inhibitory protein, mammary inhibitory activity (MIA), was purified to apparent homogeneity from human milk. At concentrations of 5 to 10 ng/ml, the factor inhibited the growth of mammary epithelial cells by 30–80% and also inhibited the growth of normal rat kidney cells. Whereas the cell division of normal human mammary epithelium in primary culture was inhibited by MIA, cell division by fibroblasts from the same tissues was unresponsive. Inhibition was dose and time dependent and readily reversed when MIA was removed. MIA also inhibited growth in culture for three cell lines. The growth inhibitory protein migrated as a 14 kDa protein under reducing conditions on polyacrylamide gels in the presence of sodium dodecyl sulfate. The apparent isoelectric point was pI 5.0. The amino acid composition of MIA resembled that of α-lactalbumin, and sequence analysis of the N-terminal region comprising residues 1–24 and an isolated peptide were identical with the N-terminal and residues 66–81 of human α-lactalbumin. In addition, MIA was active in the lactose synthase system. The results strongly suggest that MIA and α-lactalbumin are identical proteins. Consistent with these results, α-lactalbumin preparations from several mammalian species, including human, goat, cow and camel, were all found to be growth inhibitory for cultured mammary epithelial cells. The inhibitory activity associated with human α-lactalbumin was destroyed by digestion with pepsin or chymotrypsin, by carboxymethylation of cysteine, or by cleavage of methionine 90 following cyanogen bromide treatment. The results raise the possibility that during lactation α-lactalbumin, a product of mammary cell differentiation, could be a physiologically relevant feed-back inhibitor of mammary cell growth and perhaps of other cell types as well.

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Abbreviations

MIA:

mammary inhibitory activity

MDGI:

mammary derived growth inhibitor

α-LA:

alpha lactalbumin

H-α-LA:

human α-lactalbumin

NRK:

normal rat kidney

IMEM:

improved minimal essential medium

DMEM:

Dulbecco's modified Eagles medium

FCS:

fetal calf serum

EGF:

epidermal growth factor

TGF β:

transforming growth factor β

CNBr:

cyanogen bromide

SDS:

sodium dodecyl sulfate

kDa:

kilodaltons

ND-PAGE:

non-denaturing polyacrylamide gel electrophoresis

TCA:

trichloroacetic acid

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Authors and Affiliations

  1. Cancer Biology Division, School of Biological Sicences, Madurai Kamaraj University, Madurai, India

    Sanjeeva Mohanam

  2. Surgery Branch, National Cancer Institute, NIH, Bethesda, Maryland

    Sue Liu

  3. Cellco Advanced Bioreactors, Inc., Kensington, Maryland

    W. R. Kidwell

  4. Department of Cell Biology, Baylor College of Medicine, Houston, Texas

    M. P. Bansal & D. Medina

  5. Howard Hughes Medical Institute and Department of Microbiology and Immunology, Baylor College of Medicine, Houston, Texas

    R. G. Cook

  6. Department of Medicinal and Analytical Chemistry, Genentech Inc., South San Francisco, California

    Claire E. Kotts

  7. Lombardi Cancer Research Center, Georgetown University, Washington, D.C.

    Mozeena Bano

  8. ARS, Eastern Regional Research Center, USDA, 600 E. Mermaid Lane, 19118, Philadelphia, PA, USA

    M. P. Thompson & H. M. Farrell Jr.

Authors
  1. M. P. Thompson
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  2. H. M. Farrell Jr.
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  3. Sanjeeva Mohanam
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  4. Sue Liu
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  5. W. R. Kidwell
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  8. D. Medina
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  9. Claire E. Kotts
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  10. Mozeena Bano
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Thompson, M.P., Farrell, H.M., Mohanam, S. et al. Identification of human milk α-lactalbumin as a cell growth inhibitor. Protoplasma 167, 134–144 (1992). https://doi.org/10.1007/BF01403376

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  • DOI: https://doi.org/10.1007/BF01403376

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