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鉴定莱茵衣藻截短型血红蛋白 THB1：与氮代谢的关联及赖氨酸作为远端血红素配体的鉴定。

Characterization of THB1, a Chlamydomonas reinhardtii truncated hemoglobin: linkage to nitrogen metabolism and identification of lysine as the distal heme ligand.

机构信息

Department of Biophysics, Johns Hopkins University , Baltimore, Maryland 21218, United States.

出版信息

Biochemistry. 2014 Jul 22;53(28):4573-89. doi: 10.1021/bi5005206. Epub 2014 Jul 9.

DOI:10.1021/bi5005206
PMID:24964018
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4108185/
Abstract

The nuclear genome of the model organism Chlamydomonas reinhardtii contains genes for a dozen hemoglobins of the truncated lineage. Of those, THB1 is known to be expressed, but the product and its function have not yet been characterized. We present mutagenesis, optical, and nuclear magnetic resonance data for the recombinant protein and show that at pH near neutral in the absence of added ligand, THB1 coordinates the heme iron with the canonical proximal histidine and a distal lysine. In the cyanomet state, THB1 is structurally similar to other known truncated hemoglobins, particularly the heme domain of Chlamydomonas eugametos LI637, a light-induced chloroplastic hemoglobin. Recombinant THB1 is capable of binding nitric oxide (NO(•)) in either the ferric or ferrous state and has efficient NO(•) dioxygenase activity. By using different C. reinhardtii strains and growth conditions, we demonstrate that the expression of THB1 is under the control of the NIT2 regulatory gene and that the hemoglobin is linked to the nitrogen assimilation pathway.

摘要

模式生物莱茵衣藻的核基因组包含十几个截断谱系的血红蛋白基因。其中，已知 THB1 是表达的，但尚未对其产物及其功能进行表征。我们提供了重组蛋白的诱变、光学和核磁共振数据，并表明在没有添加配体的情况下，近中性 pH 值下，THB1 用典型的近端组氨酸和远端赖氨酸与血红素铁配位。在氰基态下，THB1 在结构上类似于其他已知的截断血红蛋白，特别是来自衣藻 Eugametos LI637 的光诱导质体血红蛋白的血红素结构域。重组 THB1 能够在铁或亚铁状态下结合一氧化氮（NO(•)），并且具有高效的 NO(•)双氧酶活性。通过使用不同的莱茵衣藻菌株和生长条件，我们证明 THB1 的表达受 NIT2 调节基因的控制，并且该血红蛋白与氮同化途径相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/e976e370d027/bi-2014-005206_0014.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/887302e435e0/bi-2014-005206_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/010d7f037c46/bi-2014-005206_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/3311f3d7091f/bi-2014-005206_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/ea4b04d25397/bi-2014-005206_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/c3f974c0df78/bi-2014-005206_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/f52906606734/bi-2014-005206_0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/6f03fe8089fd/bi-2014-005206_0008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/d191684025ec/bi-2014-005206_0009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/30930f5172ba/bi-2014-005206_0010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/88bfc2ccd7c7/bi-2014-005206_0011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/f723af2554fe/bi-2014-005206_0012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/8f648a69bb64/bi-2014-005206_0013.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/e976e370d027/bi-2014-005206_0014.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/887302e435e0/bi-2014-005206_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/010d7f037c46/bi-2014-005206_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/3311f3d7091f/bi-2014-005206_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/ea4b04d25397/bi-2014-005206_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/c3f974c0df78/bi-2014-005206_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/f52906606734/bi-2014-005206_0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/6f03fe8089fd/bi-2014-005206_0008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/d191684025ec/bi-2014-005206_0009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/30930f5172ba/bi-2014-005206_0010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/88bfc2ccd7c7/bi-2014-005206_0011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/f723af2554fe/bi-2014-005206_0012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/8f648a69bb64/bi-2014-005206_0013.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/838d/4108185/e976e370d027/bi-2014-005206_0014.jpg

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