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Synthesis and expression in Escherichia coli of the gene encoding monocyte-derived neutrophil-activating factor: biological equivalence between natural and recombinant neutrophil-activating factor.
Open accessAbstract
The neutrophil-activating factor (NAF) purified from the conditioned medium of lipopolysaccharide-stimulated human monocytes was sequenced and found to consist of 72 amino acids: SAKELRCQCIKTYSKPFHPKFIKELRVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRA ENS. Purified preparations of natural NAF contained, in addition to this main form, minor amounts of three amino-terminal variants with 77 (+AVLPR), 70, and 69 residues. A gene coding for the 72-amino acid NAF was synthesized, cloned, and expressed in Escherichia coli. Western (immunologic) blot analysis of crude bacterial extracts, with an antiserum raised against natural NAF, revealed a single band that comigrated with natural NAF. Recombinant NAF purified to homogeneity had identical amino- and carboxyl-terminal sequences to the 72-amino acid natural NAF. Recombinant NAF was tested on human neutrophils and had the same activity and potency as natural NAF in inducing chemotaxis, rapidly increasing cytosolic free Ca2+, activating the respiratory burst, and releasing specific and azurophilic granular contents.
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Cites
A rapid alkaline extraction procedure for screening recombinant plasmid DNA
Birnboim, H. C., & Doly, J. (1979) · Nucleic Acids Research · doi:10.1093/nar/7.6.1513 · Cited by 14,686
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Matsushima, K., & Oppenheim, J. J. (1989) · Cytokine · doi:10.1016/1043-4666(89)91043-0 · Cited by 607
Clark-Lewis, I., Schumacher, C., Baggiolini, M., & Moser, B. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)54472-0 · Cited by 425
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de Oliveira, S., Reyes-Aldasoro, C. C., Candel, S., Renshaw, S. A., Mulero, V., & Calado, Â. (2013) · The Journal of Immunology · doi:10.4049/jimmunol.1203266 · Cited by 381
Walz, A., Burgener, R., Car, B., Baggiolini, M., Kunkel, S. L., & Strieter, R. M. (1991) · The Journal of experimental medicine · doi:10.1084/jem.174.6.1355 · Cited by 353
Metadata
- Authors
- Lindley, I., Aschauer, H., Seifert, J. M., Lam, C., Brunowsky, W., Kownatzki, E., Thelen, M., Peveri, P., Dewald, B., & von Tscharner, V
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1988-12
- Volume
- 85
- Issue
- 23
- Pages
- 9199-9203
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1969904510
- Cited by
- 230
- References
- 1
- FWCI
- 8.67
- Type
- journal article
- Language
- en
- Open access
- green
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- Lindley, I., Aschauer, H., Seifert, J. M., Lam, C., Brunowsky, W., Kownatzki, E., Thelen, M., Peveri, P., Dewald, B., & von Tscharner, V. (1988). Synthesis and expression in Escherichia coli of the gene encoding monocyte-derived neutrophil-activating factor: biological equivalence between natural and recombinant neutrophil-activating factor. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.85.23.9199
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