Difference between revisions of "Part:BBa K1998012"

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		+	The wrong sequence was submitted to this page in 2016. iGEM Macquarie 2017 has created a new updated part page with the correct sequence (BBa_K2300003) (https://parts.igem.org/Part:BBa_K2300003#Overview).
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Latest revision as of 00:39, 2 November 2017

HydEF

The wrong sequence was submitted to this page in 2016. iGEM Macquarie 2017 has created a new updated part page with the correct sequence (BBa_K2300003) (https://parts.igem.org/Part:BBa_K2300003#Overview).

Sequence and Features

Overview

The HydEF protein contains two unique domains that are homologous to two distinct prokaryotic proteins, HydE and HydF, which are found exclusively in organisms containing [Fe] hydrogenase. This part makes up one of the parts found in an operon of the hydrogen production pathway.

Biology & Literature

hydEF (comprising of HydE and HydF) encodes proteins necessary for hydrogenase activity by functioning in H cluster biosynthesis and enzyme maturation involving S-adenosylmethionine (SAM) dependent mechanisms [1]. HydF is part of the GTPase protein family, with GTP-binding motifs present at the N terminal end and iron-sulfur cluster-binding motifs at the C terminal end [2]. Both HydE is a radical SAM (S-adenosyl methionine) enzyme [3]. King et al. stated that challenges faced during the expression of maturation genes HydEF and HydG in E. coli were that they were unstable and prone to rearrangements [1].

Both HydEF and HydG are transcribed under anaerobic conditions and are necessary for the assembly of an active HydA1 hydrogenase [2]. The importance of HydEF in hydrogenase activity has also been illustrated in the mutant hydEF-1 strain of C. reinhardtii which is unable to produce H2 due to a non-functional HydEF gene [4].

Part Verification

Fig 1. A gel consisting of EcorI/PstI for the hydEF part. The expected band for the hydEF (3611 bp) part was observed.

Protein information

HydEF
Mass: 121.95 kDa
Sequence:
MAHSLSAHSRQAGDRKLGAGAASSRPSCPSRRIVRVAAHASASKATPDVPVDDLPPAHARAAVAAANRRARAMASAEAAAETLGDFLGLGKGGLSP GATANLDREQVLGVLEAVWRRGDLNLERALYSHANAVTNKYCGGGVYYRGLVEFSNICQNDCSYCGIRNNQKEVWRYTMPVEEVVEVAKWALENGI RNIMLQGGELKTEQRLAYLEACVRAIREETTQLDLEMRARAASTTTAEAAASAQADAEAKRGEPELGVVVSLSVGELPMEQYERLFRAGARRYLIRIET SNPDLYAALHPEPMSWHARVECLRNLKKAGYMLGTGVMVGLPGQTLHDLAGDVMFFRDIKADMIGMGPFITQPGTPATDKWTALYPNANKNSHMK SMFDLTTAMNALVRITMGNVNISATTALQAIIPTGREIALERGANVVMPILTPTQYRESYQLYEGKPCITDTAVQCRRCLDMRLHSVGKTSAAGVWGDPA SFLHPIVGVPVPHDLSSPALAAAASADFHEVGAGPWNPIRLERLVEVPDRYPDPDNHGRKKAGAGKGGKAHDSHDDGDHDDHHHHHGAAPAGAAA GKGTGAAAIGGGAGASRQRVAGAAAASARLCAGARRAGRVVASPLRPAAACRGVAVKAAAAAAGEDAGAGTSGVGSNIVTSPGIASTTAHGVPRINI GVFGVMNAGKSTLVNALAQQEACIVDSTPGTTADVKTVLLELHALGPAKLLDTAGLDEVGGLGDKKRRKALNTLKECDVAVLVVDTDTAAAAIKSGRLA EALEWESKVMEQAHKYNVSPVLLLNVKSRGLPEAQAASMLEAVAGMLDPSKQIPRMSLDLASTPLHERSTITSAFVKEGAVRSSRYGAPLPGCLPRW SLGRNARLLMVIPMDAETPGGRLLRPQAQVMEEAIRHWATVLSVRLDLDAARGKLGPEACEMERQRFDGVIAMMERNDGPTLVVTDSQAIDVVHPW TLDRSSGRPLVPITTFSIAMAYQQNGGRLDPFVEGLEALETLQDGDRVLISEACNHNRITSACNDIGMVQIPNKLEAALGGKKLQIEHAFGREFPELESG GMDGLKLAIHCGGCMIDAQKMQQRMKDLHEAGVPVTNYGVFFSWAAWPDALRRALEPWGVEPPVGTPATPAAAPATAASGV

References

[1] King, P., Posewitz, M., Ghirardi, M. and Seibert, M. (2006). Functional Studies of [FeFe] Hydrogenase Maturation in an Escherichia coli Biosynthetic System. Journal of Bacteriology, 188(6), pp.2163-2172.

[2] Posewitz, M., King, P., Smolinski, S., Zhang, L., Seibert, M. and Ghirardi, M. (2004). Discovery of Two Novel Radical S-Adenosylmethionine Proteins Required for the Assembly of an Active [Fe] Hydrogenase. Journal of Biological Chemistry, 279(24), pp.25711-25720.

[3] Kelly, C., Pinske, C., Murphy, B., Parkin, A., Armstrong, F., Palmer, T. and Sargent, F. (2015). Integration of an [FeFe]-hydrogenase into the anaerobic metabolism of Escherichia coli. Biotechnology Reports, 8, pp.94-104.

[4] Posewitz, M., King, P., Smolinski, S., Smith, R., Ginley, A., Ghirardi, M. and Seibert, M. (2005). Identification of genes required for hydrogenase activity in Chlamydomonas reinhardtii : Figure 1. Biochm. Soc. Trans., 33(1), pp.102-104.