Role of Subunit NuoL for Proton Translocation by Respiratory Complex I

Steimle, Stefan; Bajzath, Csaba; Schulte, Marius; Bothe, Vinzenz; Friedrich, Thorsten

doi:10.1021/bi200264q

Cited by 60 publications

(51 citation statements)

References 45 publications

(89 reference statements)

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“…For example, a recent site-directed mutagenesis analysis of NuoL showed that mutation of a site that was detected by TreeSAAP and statistically coupled to the sites under selection (site #178) caused a significant reduction in Complex I activity [73], [74]. Other work has shown that removal of the piston arm also reduces Complex I activity [75]. Our results provide additional sites that can be tested for altered Complex I activity with mutational analyses and also provide methods to carry out a SCA in other taxonomic groups for other functionally important sites to empirically test in model organisms.…”

Section: Discussionmentioning

confidence: 99%

Positive Darwinian Selection in the Piston That Powers Proton Pumps in Complex I of the Mitochondria of Pacific Salmon

2011

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The mechanism of oxidative phosphorylation is well understood, but evolution of the proteins involved is not. We combined phylogenetic, genomic, and structural biology analyses to examine the evolution of twelve mitochondrial encoded proteins of closely related, yet phenotypically diverse, Pacific salmon. Two separate analyses identified the same seven positively selected sites in ND5. A strong signal was also detected at three sites of ND2. An energetic coupling analysis revealed several structures in the ND5 protein that may have co-evolved with the selected sites. These data implicate Complex I, specifically the piston arm of ND5 where it connects the proton pumps, as important in the evolution of Pacific salmon. Lastly, the lineage to Chinook experienced rapid evolution at the piston arm.

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Section: Discussionmentioning

confidence: 99%

Positive Darwinian Selection in the Piston That Powers Proton Pumps in Complex I of the Mitochondria of Pacific Salmon

2011

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“…Steimle et al (28) reported that overexpressed NDH-1 isolated from NuoL knock-out mutant of E. coli displayed 80% of piericidin-sensitive NADH-DB reductase activity compared with that of the wild type. The membrane preparations of this mutant showed approximately twice the NADH-K 3 Fe(CN) 6 reductase activity as that of the wild type.…”

Section: Discussionmentioning

confidence: 99%

Structural Contribution of C-terminal Segments of NuoL (ND5) and NuoM (ND4) Subunits of Complex I from Escherichia coli

Torres‐Bacete

Sinha

Matsuno‐Yagi

et al. 2011

Journal of Biological Chemistry

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The proton-translocating NADH-quinone oxidoreductase (complex I/NDH-1) is a multisubunit enzymatic complex. It has a characteristic L-shaped form with two domains, a hydrophilic peripheral domain and a hydrophobic membrane domain. The membrane domain contains three antiporter-like subunits (NuoL, NuoM, and NuoN, Escherichia coli naming) that are considered to be involved in the proton translocation. Deletion of either NuoL or NuoM resulted in an incomplete assembly of NDH-1 and a total loss of the NADH-quinone oxidoreductase activity. We have truncated the C terminus segments of NuoM and NuoL by introducing STOP codons at different locations using site-directed mutagenesis of chromosomal DNA. Our results suggest an important structural role for the C-terminal segments of both subunits. The data further advocate that the elimination of the last transmembrane helix (TM14) of NuoM and the TM16 (at least C-terminal seven residues) or together with the HL helix and the TM15 of the NuoL subunit lead to reduced stability of the membrane arm and therefore of the whole NDH-1 complex. A region of NuoL critical for stability of NDH-1 architecture has been discussed.

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“…Subunit L differs from the related subunits M and N in that it has 2 additional TM (transmembrane) helices, which are connected by a long lateral helix that contacts subunits M, N and K (Efremov and Sazanov 2011). The role of the lateral helix has been tested by mutagenesis in several recent studies (Belevich et al 2011; Steimle et al 2011; Steimle et al 2015; Steimle et al 2012; Torres-Bacete et al 2011). Proton translocation likely occurs through four pathways.…”

Section: Introductionmentioning

confidence: 99%

Loss of Complex I activity in the Escherichia coli enzyme results from truncating the C-terminus of subunit K, but not from cross-linking it to subunits N or L

Zhu

Canales

Bedair

et al. 2016

J Bioenerg Biomembr

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Complex I is a multi-subunit enzyme of the respiratory chain with seven core subunits in its membrane arm (A, H, J, K, L, M, and N). In the enzyme from Escherichia coli the C-terminal ten amino acids of subunit K lie along the lateral helix of subunit L, and contribute to a junction of subunits K, L and N on the cytoplasmic surface. Using double cysteine mutagenesis, the cross-linking of subunit K (R99C) to either subunit L (K581C) or subunit N (T292C) was attempted. A partial yield of cross-linked product had no effect on the activity of the enzyme, or on proton translocation, suggesting that the C-terminus of subunit K has no dynamic role in function. To further elucidate the role of subunit K genetic deletions were constructed at the C-terminus. Upon the serial deletion of the last 4 residues of the C-terminus of subunit K, various results were obtained. Deletion of one amino acid had little effect on the activity of Complex I, but deletions of 2 or more amino acids led to total loss of enzyme activity and diminished levels of subunits L, M, and N in preparations of membrane vesicles. Together these results suggest that while the C-terminus of subunit K has no dynamic role in energy transduction by Complex I, it is vital for the correct assembly of the enzyme.

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Role of Subunit NuoL for Proton Translocation by Respiratory Complex I

Cited by 60 publications

References 45 publications

Positive Darwinian Selection in the Piston That Powers Proton Pumps in Complex I of the Mitochondria of Pacific Salmon

Positive Darwinian Selection in the Piston That Powers Proton Pumps in Complex I of the Mitochondria of Pacific Salmon

Structural Contribution of C-terminal Segments of NuoL (ND5) and NuoM (ND4) Subunits of Complex I from Escherichia coli

Loss of Complex I activity in the Escherichia coli enzyme results from truncating the C-terminus of subunit K, but not from cross-linking it to subunits N or L

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