Proteolytic activity of bovine lactoferrin

Maria Teresa Massucci, Francesco Giansanti, Giovanna Di Nino, Manola Turacchio, Maria Federica Giardi, Dario Botti, Rodolfo Ippoliti, Barbara De Giulio, Rosa Siciliano, Giovanna Donnarumma, Piera Valenti, Alessio Bocedi, Fabio Polticelli, Paolo Ascenzi, Giovanni Antonini

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Bovine lactoferrin catalyzes the hydrolysis of synthetic substrates (i.e., Z-aminoacyl-7-amido-4-metliylcoumarin). Values of Km and k cat for the bovine lactoferrin catalyzed hydrolysis of Z-Phe-Arg-7-amido-4-methylcoumarin are 50 μM and 0.03 s-1, respectively, the optimum pH value is 7.5 at 25°C. The bovine lactoferrin substrate specificity is similar to that of trypsin, while the hydrolysis rate is several orders of magnitude lower than that of trypsin. The bovine lactoferrin catalytic activity is irreversibly inhibited by the serine-protease inhibitors PMSF and Pefabloc. Moreover, both iron-saturation of the protein and LPS addition strongly inhibit the bovine lactoferrin activity. Interestingly, bovine lactoferrin undergoes partial auto-proteolytic cleavage at positions Arg415-Lys 416 and Lys440-Lys441. pKa shift calculations indicate that several Ser residues of bovine lactoferrin display the high nucleophilicity required to potentially catalyze substrate cleavage. However, a definitive identification of the active site awaits further studies.

Original languageEnglish
Pages (from-to)249-255
Number of pages7
JournalBioMetals
Volume17
Issue number3
DOIs
Publication statusPublished - Jun 2004

Fingerprint

Lactoferrin
lactoferrin
Hydrolysis
cattle
Substrates
hydrolysis
Catalyst activity
Trypsin
trypsin
Iron
Proteins
Serine Proteinase Inhibitors
serine proteinases
substrate specificity
Substrate Specificity
proteinase inhibitors
catalytic activity
automobiles
active sites
Catalytic Domain

Keywords

  • Autoproteolysis
  • Iron
  • Lactoferrin
  • LPS

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Massucci, M. T., Giansanti, F., Di Nino, G., Turacchio, M., Giardi, M. F., Botti, D., ... Antonini, G. (2004). Proteolytic activity of bovine lactoferrin. BioMetals, 17(3), 249-255. https://doi.org/10.1023/B:BIOM.0000027700.90780.45

Proteolytic activity of bovine lactoferrin. / Massucci, Maria Teresa; Giansanti, Francesco; Di Nino, Giovanna; Turacchio, Manola; Giardi, Maria Federica; Botti, Dario; Ippoliti, Rodolfo; De Giulio, Barbara; Siciliano, Rosa; Donnarumma, Giovanna; Valenti, Piera; Bocedi, Alessio; Polticelli, Fabio; Ascenzi, Paolo; Antonini, Giovanni.

In: BioMetals, Vol. 17, No. 3, 06.2004, p. 249-255.

Research output: Contribution to journalArticle

Massucci, MT, Giansanti, F, Di Nino, G, Turacchio, M, Giardi, MF, Botti, D, Ippoliti, R, De Giulio, B, Siciliano, R, Donnarumma, G, Valenti, P, Bocedi, A, Polticelli, F, Ascenzi, P & Antonini, G 2004, 'Proteolytic activity of bovine lactoferrin', BioMetals, vol. 17, no. 3, pp. 249-255. https://doi.org/10.1023/B:BIOM.0000027700.90780.45
Massucci MT, Giansanti F, Di Nino G, Turacchio M, Giardi MF, Botti D et al. Proteolytic activity of bovine lactoferrin. BioMetals. 2004 Jun;17(3):249-255. https://doi.org/10.1023/B:BIOM.0000027700.90780.45
Massucci, Maria Teresa ; Giansanti, Francesco ; Di Nino, Giovanna ; Turacchio, Manola ; Giardi, Maria Federica ; Botti, Dario ; Ippoliti, Rodolfo ; De Giulio, Barbara ; Siciliano, Rosa ; Donnarumma, Giovanna ; Valenti, Piera ; Bocedi, Alessio ; Polticelli, Fabio ; Ascenzi, Paolo ; Antonini, Giovanni. / Proteolytic activity of bovine lactoferrin. In: BioMetals. 2004 ; Vol. 17, No. 3. pp. 249-255.
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