Stoichiometry of two plant glycine decarboxylase complexes and comparison with a cyanobacterial glycine cleavage system. (12th May 2020)

• Record Type: Journal Article
• Authors: Wittmiß, Maria; Mikkat, Stefan; Hagemann, Martin; Bauwe, Hermann
• Is Part Of: Plant journal. Volume 103:Number 2(2020)
• Page Start: 801
• Page End: 813
• Publication Date: 2020-05-12
• Subjects: Arabidopsis thaliana -- glycine cleavage system -- glycine decarboxylase complex -- mitochondria -- multienzyme metabolic complexes -- one‐carbon metabolism -- photorespiration -- Pisum sativum -- recombinant proteins -- Synechocystis; Plant molecular biology -- Periodicals; Plant cells and tissues -- Periodicals; Botany -- Periodicals; 580
• DOI: 10.1111/tpj.14773
• Languages: English
• ISSNs: 0960-7412

Abstract

Summary: The multienzyme glycine cleavage system (GCS) converts glycine and tetrahydrofolate to the one‐carbon compound 5, 10‐methylenetetrahydrofolate, which is of vital importance for most if not all organisms. Photorespiring plant mitochondria contain very high levels of GCS proteins organised as a fragile glycine decarboxylase complex (GDC). The aim of this study is to provide mass spectrometry‐based stoichiometric data for the plant leaf GDC and examine whether complex formation could be a general property of the GCS in photosynthesizing organisms. The molar ratios of the leaf GDC component proteins are 1L2 ‐4P2 ‐8T‐26H and 1L2 ‐4P2 ‐8T‐20H for pea and Arabidopsis, respectively, as determined by mass spectrometry. The minimum mass of the plant leaf GDC ranges from 1550 to 1650 kDa, which is larger than previously assumed. The Arabidopsis GDC contains four times more of the isoforms GCS‐P1 and GCS‐L1 in comparison with GCS‐P2 and GCS‐L2, respectively, whereas the H‐isoproteins GCS‐H1 and GCS‐H3 are fully redundant as indicated by their about equal amounts. Isoform GCS‐H2 is not present in leaf mitochondria. In the cyanobacterium Synechocystis sp. PCC 6803, GCS proteins concentrations are low but above the complex formation threshold reported for pea leaf GDC. Indeed, formation of a cyanobacterial GDC from the individual recombinant GCS proteins in vitro could be demonstrated. Presence and metabolic significance of a Synechocystis GDC in vivo remain to be examined but could involve multimers of the GCS H‐protein that dynamically crosslink the three GCS enzyme proteins, facilitating glycine metabolism by the formation of multienzyme metabolic complexes. Data are available via ProteomeXchange with identifier PXD018211. Significance Statement: This work provides first mass spectrometry‐based stoichiometric data for glycine decarboxylase complexes from two plant species in comparison with a cyanobacterial glycine cleavage system and examines whether complex formation could be a general property of this important ubiquitous multienzyme system.