Biochemical properties of the matrix metalloproteinase NtMMP1 from Nicotiana tabacum cv. BY-2 suspension cells.

Planta
Manoj K MandalAndreas Schiermeyer

Abstract

A zinc-dependent matrix metalloproteinase (NtMMP1) found in the plasma membrane of Nicotiana tabacum cv. Bright Yellow 2 (BY-2) suspension cells is thought to be responsible for the degradation of recombinant proteins secreted into the culture supernatant. We have characterized the proteolytic activity of NtMMP1 by expressing a recombinant derivative lacking the C-terminal transmembrane domain in yeast. After purifying the protein by affinity chromatography, its autocatalytic activity was analyzed using monoclonal antibodies raised against its N-terminal and C-terminal portions. Both the unprocessed and processed forms of NtMMP1 displayed caseinolytic activity and N-terminal sequencing identified an autocatalytic cleavage site within the sequence motif HFSFFP, which is similar to the corresponding sequences of the human matrix metalloproteinases stromelysin-1 (MMP-3) and stromelysin-2 (MMP-10). Unlike all other matrix metalloproteinases investigated so far, NtMMP1 contains a disulfide bond within its propeptide thus rendering the proenzyme catalytically active. Kinetic analysis of NtMMP1 with a synthetic substrate revealed a K(m) of 10.55 +/- 0.9 microM, a k(cat) of 0.6 +/- 0.01 s(-1) and maximum activity at pH 7.5. We found th...Continue Reading

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Citations

Dec 9, 2015·BMC Genetics·Chiara BroccanelloMassimo Saccomani
Mar 26, 2016·Frontiers in Plant Science·Manoj K MandalAndreas Schiermeyer
Mar 26, 2016·Frontiers in Plant Science·Rita B SantosTanja Holland
Nov 17, 2011·Physiologia Plantarum·Giada Marino, Christiane Funk
Feb 24, 2015·Frontiers in Plant Science·Jérôme LallemandPierre Tocquin
Apr 5, 2018·Plant Biotechnology Journal·Philippe V JutrasDominique Michaud
Mar 7, 2018·Plant Biotechnology Journal·Friederike Grosse-HolzRenier A L van der Hoorn
Jan 20, 2021·Journal of Experimental Botany·Alice Godson, Renier A L van der Hoorn
Apr 11, 2017·Biochimica Et Biophysica Acta. Molecular Cell Research·Laura Marino-PuertasF Xavier Gomis-Rüth
Sep 19, 2018·Journal of Proteome Research·Sebastian N W HoernsteinRalf Reski

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