Substitution of the sole tryptophan of the cupredoxin, amicyanin, with 5-hydroxytryptophan alters fluorescence properties and energy transfer to the type 1 copper site

Pastore, Anthony J.; Ficaretta, Elise D.; Chatterjee, Abhishek; Davidson, Victor L.

doi:10.1016/j.jinorgbio.2022.111895

Cited by 3 publications

(4 citation statements)

References 43 publications

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“…The incorporation of 5-HTP changed the fluorescence emission maximum of amicyanin from 318 to 331 nm. Moreover, the fluorescence quantum yield of the 5-HTP-containing amicyanin was much less than that of the native amicyanin [ 58 ]. This example illustrated that the incorporation of ncAAs changed the physical properties of proteins.…”

Section: Methods For Ncaas Incorporation Into Proteinsmentioning

confidence: 99%

Advances in Biosynthesis of Non-Canonical Amino Acids (ncAAs) and the Methods of ncAAs Incorporation into Proteins

Chen,

Xin,

Zhang

et al. 2023

Molecules

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The functional pool of canonical amino acids (cAAs) has been enriched through the emergence of non-canonical amino acids (ncAAs). NcAAs play a crucial role in the production of various pharmaceuticals. The biosynthesis of ncAAs has emerged as an alternative to traditional chemical synthesis due to its environmental friendliness and high efficiency. The breakthrough genetic code expansion (GCE) technique developed in recent years has allowed the incorporation of ncAAs into target proteins, giving them special functions and biological activities. The biosynthesis of ncAAs and their incorporation into target proteins within a single microbe has become an enticing application of such molecules. Based on that, in this study, we first review the biosynthesis methods for ncAAs and analyze the difficulties related to biosynthesis. We then summarize the GCE methods and analyze their advantages and disadvantages. Further, we review the application progress of ncAAs and anticipate the challenges and future development directions of ncAAs.

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Section: Methods For Ncaas Incorporation Into Proteinsmentioning

confidence: 99%

Advances in Biosynthesis of Non-Canonical Amino Acids (ncAAs) and the Methods of ncAAs Incorporation into Proteins

Chen,

Xin,

Zhang

et al. 2023

Molecules

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“…370 Subsequently, 5HW has been used to study other biological processes, such as proteinnucleic acid interactions 365,371 and ion binding to proteins. 367,372,373 In addition, 5HW has been used as a FRET donor to other fluorescent reporters, such as AEDANS. 374 Fluorotryptophan.…”

Section: Tryptophan-based Uaasmentioning

confidence: 99%

“…Szabo and co-workers first biosynthetically incorporated 5HW into the Ca 2+ binding protein, oncomodulin, and demonstrated the utility of 5HW as a fluorescence probe for protein–protein interactions . Subsequently, 5HW has been used to study other biological processes, such as protein-nucleic acid interactions , and ion binding to proteins. ,, In addition, 5HW has been used as a FRET donor to other fluorescent reporters, such as AEDANS …”

Section: Fluorescence Probesmentioning

confidence: 99%

Unnatural Amino Acids for Biological Spectroscopy and Microscopy

Feng,

Wang,

Zhang

et al. 2024

Chem. Rev.

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Due to advances in methods for site-specific incorporation of unnatural amino acids (UAAs) into proteins, a large number of UAAs with tailored chemical and/or physical properties have been developed and used in a wide array of biological applications. In particular, UAAs with specific spectroscopic characteristics can be used as external reporters to produce additional signals, hence increasing the information content obtainable in protein spectroscopic and/or imaging measurements. In this Review, we summarize the progress in the past two decades in the development of such UAAs and their applications in biological spectroscopy and microscopy, with a focus on UAAs that can be used as site-specific vibrational, fluorescence, electron paramagnetic resonance (EPR), or nuclear magnetic resonance (NMR) probes. Wherever applicable, we also discuss future directions.

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“…Incorporation of tyrosine analogs are well documented and have been used to probe long‐range electron transfer (LRET) in ribonucleotide reductase and cytochrome c peroxidase (Chang, Yee, et al, 2004; Ravichandran et al, 2013; Ravichandran et al, 2017; Reece & Seyedsayamdost, 2017; Yee et al, 2019). Tryptophan analogs are increasingly used in recombinant proteins, primarily for their cross‐linking ability, their fluorescence properties, their ability to serve as pH sensors, and for GCE methods development (Bae et al, 2003; Boknevitz et al, 2019; Buddha & Crane, 2005; Budisa et al, 2002; Englert et al, 2015; Hilaire et al, 2017; Italia et al, 2017; Ohler et al, 2021; Pastore et al, 2022; Pratt & Ho, 1975; Ross et al, 1992; Singh‐Blom et al, 2014; Wong & Eftink, 1997). The selective incorporation of tryptophan analogs became feasible recently by applying GCE technologies as demonstrated by Italia et al (2017), Englert et al (2015), and Ficaretta et al (2022) and is ideal for studying the biophysical role of tryptophan residues because their respective indole side chains possess different electronic properties (Ohler et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Selective incorporation of 5‐hydroxytryptophan blocks long range electron transfer in oxalate decarboxylase

et al. 2022

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Oxalate decarboxylase from Bacillus subtilis is a binuclear Mn-dependent acid stress response enzyme that converts the mono-anion of oxalic acid into formate and carbon dioxide in a redox neutral unimolecular disproportionation reaction. A π-stacked tryptophan dimer, W96 and W274, at the interface between two monomer subunits facilitates long-range electron transfer between the two Mn ions and plays an important role in the catalytic mechanism. Substitution of W96 with the unnatural amino acid 5-hydroxytryptophan leads to a persistent EPR signal which can be traced back to the neutral radical of 5-hydroxytryptophan with its hydroxyl proton removed. 5-Hydroxytryptophan acts as a hole sink preventing the formation of Mn(III) at the N-terminal active site and strongly suppresses enzymatic activity. The lower boundary of the standard reduction potential for the active site Mn(II)/Mn(III) couple can therefore be estimated as 740 mV against the normal hydrogen electrode at pH 4, the pH of maximum catalytic efficiency. Our results support the catalytic importance of long-range electron transfer in oxalate decarboxylase while at the same time highlighting the utility of unnatural amino acid incorporation and specifically the use of 5-hydroxytryptophan as an energetic sink for hole hopping to probe electron transfer in redox proteins.

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Substitution of the sole tryptophan of the cupredoxin, amicyanin, with 5-hydroxytryptophan alters fluorescence properties and energy transfer to the type 1 copper site

Cited by 3 publications

References 43 publications

Advances in Biosynthesis of Non-Canonical Amino Acids (ncAAs) and the Methods of ncAAs Incorporation into Proteins

Advances in Biosynthesis of Non-Canonical Amino Acids (ncAAs) and the Methods of ncAAs Incorporation into Proteins

Unnatural Amino Acids for Biological Spectroscopy and Microscopy

Selective incorporation of 5‐hydroxytryptophan blocks long range electron transfer in oxalate decarboxylase

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