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BioMed Research International
Volume 2013, Article ID 646539, 8 pages
Research Article

Thermal Stability of Glucokinases in Thermoanaerobacter tengcongensis

1Beijing Institute of Genomics, Chinese Academy of Sciences, No. 1 Beichen West Road, Chaoyang District, Beijing 100101, China
2Developmental Genetics Section, Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4264, USA
3Beijing Proteomics Innovation, Beijing Airport Industrial Zone B-6, Shunyi, Beijing 101318, China

Received 29 April 2013; Accepted 18 July 2013

Academic Editor: Paul W. Doetsch

Copyright © 2013 Zhong Qian et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Figure 1. Multi-sequence alignment of glucokinase candidates from T. tengcongensis compared to GLK from T. maritima. The boxed regions are possible conserved motifs.

Supplementary Figure 2. Temperature-dependence of recombinant ATP- and ADP-GLK. The enzyme-specific activities of recombinant ATP- and ADP-GLK were tested at different temperatures, from 30 to 95 °C. One hundred percent of activity corresponded to the specific activity at 75 °C (ATP-GLK) or 80 °C (ADP-GLK).

Supplementary Figure 3. Dynamic analysis of in vitro protein degradation of recombinant GLKs. Upper panel: Recombinant ATP- or ADP-GLK was incubated with (T) or without (C) T. tengcongensis native protein lysate at 80 °C for the indicated times. Then, the reactions were stopped by addition of SDS loading buffer. After SDS-PAGE separation, the remaining protein was detected by Western blot with specific anti-His6 antibody. Lower panel: the abundance of each immune band was estimated by image quantification software. The ratios between T and C were calculated and plotted, as shown.

Supplementary Figure 4. Phylogenetic analysis of glucokinases from different sources, using the maximum likelihood method. The horizontal bar represents a distance of 0.2 substitutions per site. The values above the lines are bootstrap values. Only values higher than 40 are shown. Database accession numbers or local gene tags are shown after the species names.

  1. Supplementary Material