Expanding the genetic code of mammalian cells

2017 ◽  
Vol 45 (2) ◽  
pp. 555-562 ◽  
Author(s):  
James S. Italia ◽  
Yunan Zheng ◽  
Rachel E. Kelemen ◽  
Sarah B. Erickson ◽  
Partha S. Addy ◽  
...  
Keyword(s):  
Protein Structure ◽  
Amino Acid ◽  
Mammalian Cells ◽  
Living Cells ◽  
Structure And Function ◽  
Unnatural Amino Acid ◽  
Full Potential ◽  
Recent Developments ◽  
Small Footprint ◽  
And Function

In the last two decades, unnatural amino acid (UAA) mutagenesis has emerged as a powerful new method to probe and engineer protein structure and function. This technology enables precise incorporation of a rapidly expanding repertoire of UAAs into predefined sites of a target protein expressed in living cells. Owing to the small footprint of these genetically encoded UAAs and the large variety of enabling functionalities they offer, this technology has tremendous potential for deciphering the delicate and complex biology of the mammalian cells. Over the last few years, exciting progress has been made toward expanding the toolbox of genetically encoded UAAs in mammalian cells, improving the efficiency of their incorporation and developing innovative applications. Here, we provide our perspective on these recent developments and highlight the current challenges that must be overcome to realize the full potential of this technology.

The construction of an amino acid network for understanding protein structure and function

Amino Acids ◽  
2014 ◽  
Vol 46 (6) ◽  
pp. 1419-1439 ◽  
Author(s):  
Wenying Yan ◽  
Jianhong Zhou ◽  
Maomin Sun ◽  
Jiajia Chen ◽  
Guang Hu ◽  
...  
Keyword(s):  
Protein Structure ◽  
Amino Acid ◽  
Structure And Function ◽  
Protein Structure And Function ◽  
And Function

scholarly journals Notes On Synthesis Of Perdeutero-5-13C,5,5,5-Trifluoroisoleucine VI

2017 ◽  
Author(s):  
David Naugler ◽  
Robert Scott Prosser
Keyword(s):  
Protein Structure ◽  
Amino Acid ◽  
Structure And Function ◽  
Methyl Groups ◽  
Nmr Studies ◽  
Branched Chain Amino Acid ◽  
Acid Methyl ◽  
Deuterated Proteins ◽  
Branched Chain ◽  
And Function

The 13CF3 group is a promising label for heteronuclear (19F,13C) NMR studies of proteins. Desirable locations for this NMR spin label include the branched chain amino acid methyl groups. It is known that replacement of CH3 by CF3 at such locations preserves protein structure and function and enhances stability. In particular, 13CF3 may be introduced at the δ position of isoleucine and incorporated biosynthetically in highly deuterated proteins. This paper reports our work in synthesis and purification of 5,5,5-trifluoroisoleucine, its perdeutero and 5-13C versions and of 2-13C-trifluoroacetate and its utility as a precursor for introduction of the 13CF3 group into proteins.

Dielectric Spectroscopy of Biomolecules up to 110 GHz

Frequenz ◽  
2018 ◽  
Vol 72 (3-4) ◽  
pp. 135-140 ◽  
Author(s):  
Eva-Maria Laux ◽  
Elena Ermilova ◽  
Daniel Pannwitz ◽  
Jessica Gibbons ◽  
Ralph Hölzel ◽  
...  
Keyword(s):  
Protein Structure ◽  
Amino Acid ◽  
Dielectric Properties ◽  
Dielectric Loss ◽  
Bacterial Culture ◽  
Structure And Function ◽  
Measuring System ◽  
Physicochemical Interaction ◽  
Dielectric Spectra ◽  
And Function

Abstract Radio-frequency fields in the GHz range are increasingly applied in biotechnology and medicine. In order to fully exploit both their potential and their risks detailed information about the dielectric properties of biological material is needed. For this purpose a measuring system is presented that allows the acquisition of complex dielectric spectra over 4 frequency decade up to 110 GHz. Routines for calibration and for data evaluation according to physicochemical interaction models have been developed. The frequency dependent permittivity and dielectric loss of some proteins and nucleic acids, the main classes of biomolecules, and of their sub-units have been determined. Dielectric spectra are presented for the amino acid alanine, the proteins lysozyme and haemoglobin, the nucleotides AMP and ATP, and for the plasmid pET-21, which has been produced by bacterial culture. Characterisation of a variety of biomolecules is envisaged, as is the application to studies on protein structure and function.

scholarly journals Investigation of Dopamine Receptor Structure and Function by Structure Prediction and Unnatural Amino Acid Mutagenesis

2012 ◽  
Vol 102 (3) ◽  
pp. 247a ◽  
Author(s):  
Ethan B. Van Arnam ◽  
Kristina N. McCleary ◽  
Fan Liu ◽  
Ravinder Abrol ◽  
Henry A. Lester ◽  
...  
Keyword(s):  
Amino Acid ◽  
Dopamine Receptor ◽  
Structure Prediction ◽  
Structure And Function ◽  
Unnatural Amino Acid ◽  
Receptor Structure ◽  
Unnatural Amino Acid Mutagenesis ◽  
Amino Acid Mutagenesis ◽  
And Function

Biological Products: Molecular Structure and Function

2021 ◽  
pp. 130-157
Keyword(s):  
Mammalian Cells ◽  
Living Cells ◽  
Structure And Function ◽  
Peptide Bonds ◽  
Biological Products ◽  
Whole Cells ◽  
Drug Products ◽  
Small Molecule Drugs ◽  
And Function ◽  
Molecular Structure And Function

This chapter analyses biological products that are defined by the method of manufacture and distinguished by a production process that separates biological drug products from orthodox, ‘small molecule’ drugs. It explains how biological products are synthesised by a variety of living cells, such as bacteria, fungi, or mammalian cells. It also refers to the types of modern biological therapies that range among proteins, nucleic acids, and whole cells. The chapter discusses monoclonal antibodies, which is considered the largest class of biological products in clinical use and are designed to attach to diseased cells, like cancer cells, that are expressing abnormal proteins on their surface. It describes proteins as chains of amino acids linked to each other chemically by peptide bonds, hence the alternative term ‘polypeptides’.

scholarly journals Structural and Functional Changes in the Na+/H+ Exchanger Isoform 1, Induced by Erk1/2 Phosphorylation

2019 ◽  
Vol 20 (10) ◽  
pp. 2378 ◽  
Author(s):  
Larry Fliegel
Keyword(s):  
Amino Acid ◽  
Mammalian Cells ◽  
Ph Regulation ◽  
Specific Binding ◽  
Structure And Function ◽  
Membrane Domain ◽  
Functional Changes ◽  
Cytosolic Domain ◽  
And Function ◽  
Disordered Regions

The human Na+/H+ exchanger isoform 1 (NHE1) is a plasma membrane transport protein that plays an important role in pH regulation in mammalian cells. Because of the generation of protons by intermediary metabolism as well as the negative membrane potential, protons accumulate within the cytosol. Extracellular signal-regulated kinase (ERK)-mediated regulation of NHE1 is important in several human pathologies including in the myocardium in heart disease, as well as in breast cancer as a trigger for growth and metastasis. NHE1 has a N-terminal, a 500 amino acid membrane domain, and a C-terminal 315 amino acid cytosolic domain. The C-terminal domain regulates the membrane domain and its effects on transport are modified by protein binding and phosphorylation. Here, we discuss the physiological regulation of NHE1 by ERK, with an emphasis on the critical effects on structure and function. ERK binds directly to the cytosolic domain at specific binding domains. ERK also phosphorylates NHE1 directly at multiple sites, which enhance NHE1 activity with subsequent downstream physiological effects. The NHE1 cytosolic regulatory tail possesses both ordered and disordered regions, and the disordered regions are stabilized by ERK-mediated phosphorylation at a phosphorylation motif. Overall, ERK pathway mediated phosphorylation modulates the NHE1 tail, and affects the activity, structure, and function of this membrane protein.

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