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Quantitative contribution of the acid production to the intracellular acidification in human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine

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Abstract

A chemotactic peptide, N-formyl-methionyl-leucyl-phenylalanine (fMLP), induced an acidification of cytosol by about 0.05 pH units in 30 sec followed by an alkalinization in human neutrophils. The quantitative contribution of acid production to the acidification was studied. The superoxide (O2 −) production stimulated by fMLP was not involved in the acidification because the production of acids in neutrophils from patients with chronic granulomatous disease who do not produce O2 −, was the same as that in normal neutrophils. The intracellular acidification was completely inhibited by deoxyglucose, suggesting that energy metabolism enhanced upon stimulation by fMLP might be the main source of the acidification. Although enhancement of the lactate formation by fMLP was 0.8 nmol/106 cells, which could lower intracellular pH by 0.08 pH units, the lactate production could not explain the initial acidification because the production of lactate started at 1 min after the stimulation while the intracellular acidification began immediately after the stimulation. Mitochondrial respiratory inhibitors such as KCN and rotenone had no effects on the fMLP-induced intracellular acidification. The fMLP-induced production of CO2 in 30 sec through the hexose monophosphate shunt was only 2.6 pmol/106 cells, which was calculated to decrease intracellular pH by only 0.0014. Thus, changes of energy metabolism induced by fMLP does not explain the acidification.

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Abbreviations

fMLP:

N-formyl-methionyl-leucyl-phenylalanine

BCECF-AM:

2′,7′-bis(carboxyethyl)carboxyfluorescein acetoxymethyl ester

PMA:

phorbol 12-myristate 13-acetate

CGD:

chronic granulomatous disease

HMP:

hexose monophosphate

pHi:

intracellular pH

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Author notes

  1. Shigeki Minakami

    Present address: Graduate School of Health and Nutrition Sciences, Nakamura Gakuen College, Befu 5-7-1, Jonan-ku, 814-01, Fukuoka, Japan

Authors and Affiliations

  1. Department of Biochemistry, Kyushu University School of Medicine, Maidashi 3-1-1, Higashi-ku, 812-82, Fukuoka, Japan

    Masaaki Satoh, Hiroichi Asagami, Dongchon Kang, Shigeki Minakami & Koichiro Takeshige

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  1. Masaaki Satoh
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  2. Hiroichi Asagami
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  3. Dongchon Kang
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  4. Shigeki Minakami
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  5. Koichiro Takeshige
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Satoh, M., Asagami, H., Kang, D. et al. Quantitative contribution of the acid production to the intracellular acidification in human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine. Mol Cell Biochem 152, 159–165 (1995). https://doi.org/10.1007/BF01076078

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  • DOI: https://doi.org/10.1007/BF01076078

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