Structural rearrangements in active and inactive forms of hydrogenase from Thiocapsa roseopersicina

K. L. Kovacs, Gabor Tigyi, L. T. Thanh, S. Lakatos, Z. Kiss, C. Bagyinka

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The electrophoretic behavior of Thiocapsa roseopersicina hydrogenase on sodium dodecyl sulfate gels demonstrates that the protein exists in two active forms, A1 and A2, which may be interconverted. Each of these forms has a characteristic electrophoretic mobility and differs in its sensitivity to O2. Form A1 is O2-labile and converts to A2 under O2. Form A2 is less sensitive to O2 and may be converted into A1 under H2 atmosphere. Both active forms are present in aerobically isolated samples. Because the proteins are still active on 1 5 % sodium dodecyl sulfate gels, they are not completely denatured, and the apparent molecular masses do not necessarily represent the true molecular masses of the enzymes. A1 has an R(f) = 0.19, corresponding to an apparent molecular mass of 90 kDa, and A2 has an R(f) = 0.35, corresponding to an apparent molecular mass of 49 k.Da. A sedimentation equilibrium centrifugation study of the active enzyme shows that the holoenzyme has a molecular mass of 98 kDa. Form A2 may be separated into two subunits of molecular mass of 64 kDa and 3 4 kDa, respectively. Thus, form A2 represents the holoenzyme with a true molecular mass of 98 kDa. Amino acid compositions and N-terminal amino acid sequences of the A2 protein and these subunits are consistent with a heterodimeric holoenzyme. The relationship between the conformational changes detected in this study and a three-state scheme proposed on the basis of FPR spectroscopic studies of the metal-containing cofactors present in the enzyme is also discussed.

Original languageEnglish (US)
Pages (from-to)947-951
Number of pages5
JournalJournal of Biological Chemistry
Volume266
Issue number2
StatePublished - Feb 19 1991
Externally publishedYes

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Thiocapsa roseopersicina
Hydrogenase
Molecular mass
varespladib methyl
Holoenzymes
Sodium Dodecyl Sulfate
Enzymes
Gels
Amino Acids
Electrophoretic mobility
Centrifugation
Protein Subunits
Atmosphere
Sedimentation
Amino Acid Sequence
Proteins
Metals

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kovacs, K. L., Tigyi, G., Thanh, L. T., Lakatos, S., Kiss, Z., & Bagyinka, C. (1991). Structural rearrangements in active and inactive forms of hydrogenase from Thiocapsa roseopersicina. Journal of Biological Chemistry, 266(2), 947-951.

Structural rearrangements in active and inactive forms of hydrogenase from Thiocapsa roseopersicina. / Kovacs, K. L.; Tigyi, Gabor; Thanh, L. T.; Lakatos, S.; Kiss, Z.; Bagyinka, C.

In: Journal of Biological Chemistry, Vol. 266, No. 2, 19.02.1991, p. 947-951.

Research output: Contribution to journalArticle

Kovacs, KL, Tigyi, G, Thanh, LT, Lakatos, S, Kiss, Z & Bagyinka, C 1991, 'Structural rearrangements in active and inactive forms of hydrogenase from Thiocapsa roseopersicina', Journal of Biological Chemistry, vol. 266, no. 2, pp. 947-951.
Kovacs, K. L. ; Tigyi, Gabor ; Thanh, L. T. ; Lakatos, S. ; Kiss, Z. ; Bagyinka, C. / Structural rearrangements in active and inactive forms of hydrogenase from Thiocapsa roseopersicina. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 2. pp. 947-951.
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