DOCSLIB.ORG

Single-Site Glycine-Specific Labeling of Proteins

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Single-Site Glycine-Specific Labeling of Proteins ARTICLE https://doi.org/10.1038/s41467-019-10503-7 OPEN Single-site glycine-specific labeling of proteins Landa Purushottam1, Srinivasa Rao Adusumalli1, Usha Singh2, V.B. Unnikrishnan1, Dattatraya Gautam Rawale1, Mansi Gujrati2, Ram Kumar Mishra2 & Vishal Rai 1 Labeling of native proteins invites interest from diverse segments of science. However, there remains the significant unmet challenge in precise labeling at a single site of a protein. Here, we report the site-specific labeling of natural or easy-to-engineer N-terminus Gly in proteins with remarkable efficiency and selectivity. The method generates a latent nucleophile from 1234567890():,; N-terminus imine that reacts with an aldehyde to deliver an aminoalcohol under physiological conditions. It differentiates N-Gly as a unique target amongst other proteinogenic amino acids. The method allows single-site labeling of proteins in isolated form and extends to lysed cells. It administers an orthogonal aldehyde group primed for late-stage tagging with an affinity tag, 19F NMR probe, and a fluorophore. A user-friendly protocol delivers analytically pure tagged proteins. The mild reaction conditions do not alter the structure and function of the protein. The cellular uptake of fluorophore-tagged insulin and its ability to activate the insulin-receptor mediated signaling remains unperturbed. 1 Department of Chemistry, Indian Institute of Science Education and Research (IISER) Bhopal, Bhopal 462066, India. 2 Department of Biological Sciences, Indian Institute of Science Education and Research (IISER) Bhopal, Bhopal 462066, India. Correspondence and requests for materials should be addressed to V.R. (email: [email protected]) NATURE COMMUNICATIONS | (2019) 10:2539 | https://doi.org/10.1038/s41467-019-10503-7 | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | https://doi.org/10.1038/s41467-019-10503-7 ingle-site labeling of proteins facilitates investigation of sev- targeting of N-terminus Gly poses a prominent complexity as Seral biological processes through attachment of biophysical there is no side-chain residue to assist. probes, imaging probes, and toxins. Such labeling emerges Here, we demonstrate that an aldehyde with appropriately through pre-engineered protein equipped with unnatural amino designed hydrogen bond acceptor can result in the exclusive acids1, an amino acid sequence recognized by enzymes2, and single labeling of N-terminus Gly residue. The remarkable selectivity of cysteine3,4 or a cysteine placed in a π-clamp5. The understanding C–C bond formation extends from an amino acid to structurally of organic chemistry with proteins has nurtured the growth of diverse proteins. Besides, the protocol operates conveniently in a bioconjugation. These biomolecules can be perceived as a macro- mixture of proteins or cell lysate. The residue-specific installation molecular substrate with multiple nucleophilic functional groups of an orthogonal aldehyde group renders analytically pure tagged (NuP). The obvious route for bioconjugation would involve its proteins in high yields through the late-stage single-site intro- reaction with an electrophile (Fig. 1a). For single-site labeling in duction of the desired probe. such cases, a functional group has to compete with all the other nucleophilic residues (chemoselectivity) and its additional copies Results (site-selectivity). The chemoselective labeling of low-frequency residues (Cys6,7,Tyr8, and Trp9) can address the latter to some We hypothesized that a multi-step transformation could provide extent. For other cases, the single-site labeling of side chain func- a roadmap to address the challenge of chemoselectivity and tionalities could be driven through ligand–protein interaction10, residue selectivity. The initial generation of the EL with capability linchpin directed modification11 or targeting the ε-amine of most to generate another reactive center (latent nucleophile, NuL, path ε 12–16 d) could provide the platform. While the first step can address reactive Lys residue (N -NH2) . Alongside, the N-terminus α α chemoselectivity, the latter can give us an opportunity to explore -amine (N -NH2) has established its place as a notable reactivity fi hotspot. In a remarkable example, the biomimetic transformation the residue speci city. Also, the NuL generated with a protein will be required to display orthogonal reactivity in the presence of of N-terminus residue delivers a carbonyl group primed for sub- fi sequent orthogonal reactions17. proteinogenic nucleophiles (Fig. 1c). At rst, the challenge was to α design an imine that can generate the NuL under physiological The kinetic labeling of N -NH2 can be achieved through its chemoselective modification with an electrophile as determined conditions. We hypothesized that H-bond acceptors in the imine by a few groups including us (Fig. 1a)18–30. The alternate route group (Fig. 1b) could assist proton shuttling and stabilize the NuL. involves a latent electrophile (E ) generated in the form of imine We anticipated that the intramolecular H-bond acceptors might α L overcome the competing intermolecular interactions with protein from N -NH2 and aldehyde (Fig. 1b). It gives us the opportunity to address the two challenges in two separate steps. If we can and solvent. Subsequently, we can explore the reactivity of NuL generate E chemoselectively, the nucleophile is only required to towards an external electrophile (Fig. 1c). Also, we anticipated L that the unsubstituted amino acid (Gly) could potentially render address the site-selectivity. This approach delivers reductive fi alkylation in a chemoselective transformation (path a, Fig. 1b)31. a reactivity pro le different from all the other substituted However, the site-selectivity gets compromised very early in the amino acids. reaction in the presence of competing imines. On the contrary, the nucleophilic attack of backbone amide to imine generated by Aminoalcohol formation with Gly under ambient conditions. α N -NH2 and 2-pyridinecarboxaldehyde can render isolable imi- We selected ortho-substituted benzaldehyde to garner the dazolidinone with high selectivity (path b, Fig. 1b)32. Unfortu- advantage of geometrical constraint imposed by the aromatic nately, none of these methodologies can distinguish one N- ring. We placed one H-bond acceptor ortho to the aldehyde in the terminus residue from the other. In this perspective, N-terminal form of a methoxy group (2a, Fig. 2a). The reaction of Gly amide Cys containing proteins can render unique reactivity to form 1a with reagent 2a fails to result in any adduct (Fig. 2a). Next, we thiazolidine with an aldehyde or 2-cyanobenzothiazole (path c, designed the reagent 2b that proffers a carbonyl group as H-bond – Fig. 1b)33 35. However, it is a challenge to identify unique reac- acceptor. It was exciting to note the formation of isolable product tivity for other N-terminus residues. In particular, selective (3ab, 27% conversion, LC-MS). Encouraged by this result, we a Selectivity landscape in labeling of 'Nup' in proteins E R H E E Reaction H Nup N Nu Nup H N p Protein: nucleophile (Nu ) 2 Protein H N Protein Nu p 2 p E O E External: electrophile (E) Nup Nup Nup E E E R′ – CHO b N-terminus labeling of proteins enabled by 'EL' Path c Path d Nup Nu R H Nu p Reaction E H p L N EL Protein Nup R′ Protein: latent electrophile (EL) N Protein Nu Nu Nu p Nu p O p External/internal: nucleophile (Nu) Nu Path b Path a Path d c This work: 'NuL' enables residue-specific labeling (R = H) E Nu p Nu Reaction R Nup p H N NuL Protein Nup Protein: latent nucleophile (Nu ) R′ L N Protein E Nu Nu External: electrophile (E) p Nu p O p NuL H Fig. 1 Residue-specificity in labeling of proteins. a Nucleophilic addition of protein residues to an electrophile. b A latent electrophile, imine, renders the N-terminus labeling of proteins (path a–path c). c Latent nucleophile enables single-site N-terminus Gly labeling 2 NATURE COMMUNICATIONS | (2019) 10:2539 | https://doi.org/10.1038/s41467-019-10503-7 | www.nature.com/naturecommunications NATURE COMMUNICATIONS | https://doi.org/10.1038/s41467-019-10503-7 ARTICLE a R OH OH CHO O O n 2 (50 mM) O O NaHCO buffer O 3 R (0.1 M, pH 7.8) NH2 CHO CHO CHO H2N ° NH2 5 mM, 25 C, 24 h NH2 2a 2b n = 3 (2c) (3ac, 33%) 1a (3aa, 0%) (3ab, 27%) n = 2 (2d) (3ad, 63%) OH O n = 1 (2e) (3a, > 95%) 3 b OH OH OH O O O O O NaHCO3 buffer O O (0.1 M, pH 7.8) 1 O H2N + R NH2 + NH2 5 mM, 25 °C, 24 h NH2 N R1 CHO NH2 Amino acid 1 2e (50 mM) OH O R1 3 4 Amide derivative of amino acids: R1 = H (3a) > 95% < 5% Gly (1a), Ala (1b), Arg (1c), Asn (1d), Asp (1e) R1 ≠ H (3b–3t) 0% Variable/reversible Cys (1f), Glu (1g), Gln (1h), His (1i), llel (1j) Leu (1k), Lys (1I), Met (1m), Phe (1n), Prol (1o) Ser (1p), Thr (1q), Trp (1r), Thr (1s), Val (1t) c O 2e, NaHCO3 buffer O O (0.1 M, pH 7.8) H2N H N H2N NH 2 NH + NH 2 5 mM, 25 °C, 24 h 2 2 Ph Ph Ph 1n 100% 0% d NuL OO OO HO NH2 HO H NH2 R1 = H R1 ≠ H 1 –H2O O N R 1a 1b–1t O N 1 + 2e O EL O O HO O O NH2 2e OH H2O 3a + 2e O N OH O O OH Fig. 2 Stable aminoalcohol formation with glycine. a Aldehyde with H-bond acceptors forms a stable aminoalcohol with 1a (also see Supplementary Fig. 72). b Selective formation of aminoalcohol with 1a. c Stereo-stability of substituted amino acid (1n) under the reaction conditions. d Plausible mechanism for glycine specific formation of aminoalcohol investigated the effect of two oxygen centers as H-bond acceptors remained inert towards any subsequent transformation contrary in the reagent 2c.
Load more

Recommended publications
  • Amino Acid Recognition by Aminoacyl-Trna Synthetases
    www.nature.com/scientificreports OPEN The structural basis of the genetic code: amino acid recognition by aminoacyl‑tRNA synthetases Florian Kaiser1,2,4*, Sarah Krautwurst3,4, Sebastian Salentin1, V. Joachim Haupt1,2, Christoph Leberecht3, Sebastian Bittrich3, Dirk Labudde3 & Michael Schroeder1 Storage and directed transfer of information is the key requirement for the development of life. Yet any information stored on our genes is useless without its correct interpretation. The genetic code defnes the rule set to decode this information. Aminoacyl-tRNA synthetases are at the heart of this process. We extensively characterize how these enzymes distinguish all natural amino acids based on the computational analysis of crystallographic structure data. The results of this meta-analysis show that the correct read-out of genetic information is a delicate interplay between the composition of the binding site, non-covalent interactions, error correction mechanisms, and steric efects. One of the most profound open questions in biology is how the genetic code was established. While proteins are encoded by nucleic acid blueprints, decoding this information in turn requires proteins. Te emergence of this self-referencing system poses a chicken-or-egg dilemma and its origin is still heavily debated 1,2. Aminoacyl-tRNA synthetases (aaRSs) implement the correct assignment of amino acids to their codons and are thus inherently connected to the emergence of genetic coding. Tese enzymes link tRNA molecules with their amino acid cargo and are consequently vital for protein biosynthesis. Beside the correct recognition of tRNA features3, highly specifc non-covalent interactions in the binding sites of aaRSs are required to correctly detect the designated amino acid4–7 and to prevent errors in biosynthesis5,8.
    [Show full text]
  • Amino Acid Chemistry
    Handout 4 Amino Acid and Protein Chemistry ANSC 619 PHYSIOLOGICAL CHEMISTRY OF LIVESTOCK SPECIES Amino Acid Chemistry I. Chemistry of amino acids A. General amino acid structure + HN3- 1. All amino acids are carboxylic acids, i.e., they have a –COOH group at the #1 carbon. 2. All amino acids contain an amino group at the #2 carbon (may amino acids have a second amino group). 3. All amino acids are zwitterions – they contain both positive and negative charges at physiological pH. II. Essential and nonessential amino acids A. Nonessential amino acids: can make the carbon skeleton 1. From glycolysis. 2. From the TCA cycle. B. Nonessential if it can be made from an essential amino acid. 1. Amino acid "sparing". 2. May still be essential under some conditions. C. Essential amino acids 1. Branched chain amino acids (isoleucine, leucine and valine) 2. Lysine 3. Methionine 4. Phenyalanine 5. Threonine 6. Tryptophan 1 Handout 4 Amino Acid and Protein Chemistry D. Essential during rapid growth or for optimal health 1. Arginine 2. Histidine E. Nonessential amino acids 1. Alanine (from pyruvate) 2. Aspartate, asparagine (from oxaloacetate) 3. Cysteine (from serine and methionine) 4. Glutamate, glutamine (from α-ketoglutarate) 5. Glycine (from serine) 6. Proline (from glutamate) 7. Serine (from 3-phosphoglycerate) 8. Tyrosine (from phenylalanine) E. Nonessential and not required for protein synthesis 1. Hydroxyproline (made postranslationally from proline) 2. Hydroxylysine (made postranslationally from lysine) III. Acidic, basic, polar, and hydrophobic amino acids A. Acidic amino acids: amino acids that can donate a hydrogen ion (proton) and thereby decrease pH in an aqueous solution 1.
    [Show full text]
  • Stimulation Effects of Foliar Applied Glycine and Glutamine Amino Acids
    Open Agriculture. 2019; 4: 164–172 Research Article Yaghoub Aghaye Noroozlo, Mohammad Kazem Souri*, Mojtaba Delshad Stimulation Effects of Foliar Applied Glycine and Glutamine Amino Acids on Lettuce Growth https://doi.org/10.1515/opag-2019-0016 received June 27, 2018; accepted January 20, 2019 1 Introduction Abstract: Amino acids have various roles in plant In biology, amino acids have vital roles in cell life. Amino metabolism, and exogenous application of amino acids acids are among the most important primary metabolites may have benefits and stimulation effects on plant growth within the plant cells. However, they are frequently and quality. In this study, the growth and nutrient uptake regarded as secondary metabolites, particularly in the of Romain lettuce (Lactuca sativa subvar Sahara) were case of proline, glycine and betaine amino acids. Many evaluated under spray of glycine or glutamine at different physiochemical characteristics of plant cells, tissues and concentrations of 0 (as control), 250, 500 and 1000 organs are influenced by the presence of amino acids (Rai mg.L-1, as well as a treatment of 250 mg.L-1 glycine+250 2002; Marschner 2011). They are the building units of mg.L-1 glutamine. The results showed that there was proteins, as the main component of living cells that have significant increase in leaf total chlorophyll content under vital roles in many cell metabolic reactions (Kielland 1994; Gly250+Glu250, Gly250 and Glu1000 mg.L-1treatments, and Rainbird et al. 1984; Jones and Darrah 1993). In addition, in leaf carotenoids content under 250 mg.L-1 glutamine amino acids have various important biological functions spray compared with the control plants.
    [Show full text]
  • Nucleotide Base Coding and Am1ino Acid Replacemients in Proteins* by Emil L
    VOL. 48, 1962 BIOCHEMISTRY: E. L. SAIITH 677 18 Britten, R. J., and R. B. Roberts, Science, 131, 32 (1960). '9 Crestfield, A. M., K. C. Smith, and F. WV. Allen, J. Biol. Chem., 216, 185 (1955). 20 Gamow, G., Nature, 173, 318 (1954). 21 Brenner, S., these PROCEEDINGS, 43, 687 (1957). 22 Nirenberg, M. WV., J. H. Matthaei, and 0. WV. Jones, unpublished data. 23 Crick, F. H. C., L. Barnett, S. Brenner, and R. J. Watts-Tobin, Nature, 192, 1227 (1961). 24 Levene, P. A., and R. S. Tipson, J. Biol. Ch-nn., 111, 313 (1935). 25 Gierer, A., and K. W. Mundry, Nature, 182, 1437 (1958). 2' Tsugita, A., and H. Fraenkel-Conrat, J. Mllot. Biol., in press. 27 Tsugita, A., and H. Fraenkel-Conrat, personal communication. 28 Wittmann, H. G., Naturwissenschaften, 48, 729 (1961). 29 Freese, E., in Structure and Function of Genetic Elements, Brookhaven Symposia in Biology, no. 12 (1959), p. 63. NUCLEOTIDE BASE CODING AND AM1INO ACID REPLACEMIENTS IN PROTEINS* BY EMIL L. SMITHt LABORATORY FOR STUDY OF HEREDITARY AND METABOLIC DISORDERS AND THE DEPARTMENTS OF BIOLOGICAL CHEMISTRY AND MEDICINE, UNIVERSITY OF UTAH COLLEGE OF MEDICINE Communicated by Severo Ochoa, February 14, 1962 The problem of which bases of messenger or template RNA' specify the coding of amino acids in proteins has been largely elucidated by the use of synthetic polyri- bonucleotides.2-7 For these triplet nucleotide compositions (Table 1), it is of in- terest to examine some of the presently known cases of amino acid substitutions in polypeptides or proteins of known structure.
    [Show full text]
  • Feedback Inhibition of Fully Unadenylylated Glutamine
    Proc. Natl. Acad. Sci. USA Vol. 90, pp. 4996-5000, June 1993 Biochemistry Feedback inhibition of fully unadenylylated glutamine synthetase from Salmonella typhimurium by glycine, alanine, and serine (x-ray crystaflography/protein structure/effector binding) SHWU-HUEY LIAW, CLARK PAN, AND DAVID EISENBERG* Molecular Biology Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90024 Contributed by David Eisenberg, December 28, 1992 ABSTRACT Bacterial glutamine synthetase (GS; EC the binding of L-glutamate to GS was measured in the 6.3.1.2) was previously shown to be inhibited by nine end presence of ADP and Pi (6, 9). Citing these studies and products of glutamine metabolism. Here we present four unpublished data, Stadtman and Ginsburg (2) concluded crystal structures ofGS, complexed with the substrate Glu and "there are separate sites on the enzyme for alanine, tryp- with each of three feedback inhibitors. The GS of the present tophan, histidine, AMP, and CTP, whereas mutually exclu- study is from Salmonela typhimurium, with Mn2+ ions bound, sive binding occurs between glycine, serine, and alanine" and is fully unadenylylated. From Fourier difference maps, we (P. Z. Smymiotis and E. R. Stadtman, unpublished data find that L-serine, L-alanine, and glycine bind at the site of the cited in review reference 2). And Rhee et al. (11) reviewed substrate L-glutamate. In our model, these four amino acids further evidence for separate sites of inhibition. In short, bind with the atoms they share in common (the "main chain" feedback inhibition ofGS is a complicated regulatory system, +NH3-CH-COO-) in the same positions.
    [Show full text]
  • Effects of Glycine on Collagen, PDGF, and EGF Expression in Model of Oral Mucositis
    nutrients Article Effects of Glycine on Collagen, PDGF, and EGF Expression in Model of Oral Mucositis Odara Maria de Sousa Sá 1,* , Nilza Nelly Fontana Lopes 2, Maria Teresa Seixas Alves 3 and Eliana Maria Monteiro Caran 4 1 Department of Pediatrics, Federal University of São Paulo, São Paulo 04023-062, Brazil 2 Former Head Division of Dentistry, Pediatric Oncology Institute, São Paulo 04023-062, Brazil; nnfl[email protected] 3 Department of Pathology, Federal University of São Paulo, São Paulo 04023-062, Brazil; [email protected] 4 Department of Pediatrics, IOP/GRAACC Medical School of Federal University of São Paulo, São Paulo 04023-062, Brazil; [email protected] * Correspondence: [email protected]; Tel.: +55-086-99932-5493 Received: 10 July 2018; Accepted: 5 September 2018; Published: 12 October 2018 Abstract: Oral mucositis is frequently a toxic effect of chemotherapeutic and/or radiotherapeutic treatment, resulting from complex multifaceted biological events involving DNA damage. The clinical manifestations have a negative impact on the life quality of cancer patients. Preventive measures and curative treatment of mucositis are still not well established. The glycine has anti-inflammatory, immunomodulatory, and cytoprotective actions, being a potential therapeutic in mucositis. The objective was to evaluate the effects of glycine on the expression of collagen and growth factors, platelet and epidermal in a hamster model oral mucositis. The mucositis was induced by the protocol of Sonis. There were 40 hamsters used, divided into two groups: Group I-control; Group II-supplemented with 5% intraperitoneal glycine, 2.0 mg/g diluted in hepes. Histopathological sections were used to perform the immune-histochemical method, the evaluation of collagen expression, and the growth factors: Epidermal growth factor (EGF) and platelet (PDGF).
    [Show full text]
  • A Study of the Renal Tubular Absorption and Glycine and Histidine in Dogs Following the Intravenous Infusion of Concetrated Solutions of These Amino Acids Daniel M
    Yale University EliScholar – A Digital Platform for Scholarly Publishing at Yale Yale Medicine Thesis Digital Library School of Medicine 1960 A study of the renal tubular absorption and glycine and histidine in dogs following the intravenous infusion of concetrated solutions of these amino acids Daniel M. Jones Yale University Follow this and additional works at: http://elischolar.library.yale.edu/ymtdl Recommended Citation Jones, Daniel M., "A study of the renal tubular absorption and glycine and histidine in dogs following the intravenous infusion of concetrated solutions of these amino acids" (1960). Yale Medicine Thesis Digital Library. 2755. http://elischolar.library.yale.edu/ymtdl/2755 This Open Access Thesis is brought to you for free and open access by the School of Medicine at EliScholar – A Digital Platform for Scholarly Publishing at Yale. It has been accepted for inclusion in Yale Medicine Thesis Digital Library by an authorized administrator of EliScholar – A Digital Platform for Scholarly Publishing at Yale. For more information, please contact [email protected]. YALE MEDICAL LIBRARY Manuscript Theses Unpublished theses submitted for the Master's and Doctor's degrees and deposited in the Yale Medical Library are to be used only with due regard to the rights of the authors. Bibliographical references may be noted, but passages must not be copied without permission of the authors, and without proper credit being given in subsequent written or published work. This thesis by has been used by the following persons, whose signatures attest their acceptance of the above restrictions. NAME AND ADDRESS DATE Digitized by the Internet Archive in 2017 with funding from The National Endowment for the Humanities and the Arcadia Fund https://archive.org/details/studyofrenaltubuOOjone A STUDY OF THE RENAL TUBULAR ABSORPTION AND EXCRETION OF GLYCINE AND HISTIDINE IN DOGS FOLLOWING THE INTRAVENOUS INFUSION OF CONCENTRATED SOLUTIONS OF THESE AMINO ACIDS DANIEL M.
    [Show full text]
  • Amino Acid Degradation
    BI/CH 422/622 OUTLINE: OUTLINE: Protein Degradation (Catabolism) Digestion Amino-Acid Degradation Inside of cells Protein turnover Dealing with the carbon Ubiquitin Fates of the 29 Activation-E1 Seven Families Conjugation-E2 nitrogen atoms in 20 1. ADENQ Ligation-E3 AA: Proteosome 2. RPH 9 ammonia oxidase Amino-Acid Degradation 18 transamination Ammonia 2 urea one-carbon metabolism free transamination-mechanism to know THF Urea Cycle – dealing with the nitrogen SAM 5 Steps Carbamoyl-phosphate synthetase 3. GSC Ornithine transcarbamylase PLP uses Arginino-succinate synthetase Arginino-succinase 4. MT – one carbon metabolism Arginase 5. FY – oxidase vs oxygenase Energetics Urea Bi-cycle 6. KW – Urea Cycle – dealing with the nitrogen 7. BCAA – VIL Feeding the Urea Cycle Glucose-Alanine Cycle Convergence with Fatty acid-odd chain Free Ammonia Overview Glutamine Glutamate dehydrogenase Overall energetics Amino Acid A. Concepts 1. ConvergentDegradation 2. ketogenic/glucogenic 3. Reactions seen before The SEVEN (7) Families B. Transaminase (A,D,E) / Deaminase (Q,N) Family C. Related to biosynthesis (R,P,H; C,G,S; M,T) 1.Glu Family a. Introduce oxidases/oxygenases b. Introduce one-carbon metabolism (1C) 2.Pyruvate Family a. PLP reactions 3. a-Ketobutyric Family (M,T) a. 1-C metabolism D. Dedicated 1. Aromatic Family (F,Y) a. oxidases/oxygenases 2. a-Ketoadipic Family (K,W) 3. Branched-chain Family (V,I,L) E. Convergence with Fatty Acids: propionyl-CoA 29 N 1 Amino Acid Degradation • Intermediates of the central metabolic pathway • Some amino acids result in more than one intermediate. • Ketogenic amino acids can be converted to ketone bodies.
    [Show full text]
  • Metabolic Classification of the Amino Acids
    Metabolic Classification of the Amino Acids *Essential and Non -essential * Glucogenic and Ketogenic 1 Essential Amino Acids • Of the 20 amino acids that make up proteins 10 of them can be synthesized by the human body • The other 10 amino acids must be acquired from food sources. These amino acids are known as essential amino acids 2 Essential Amino Acids Complete protein Incomplete protein • Contains all 10 Lack one of more of the essential amino acids essential amino acids • Proteins derived from Most vegetable proteins animal sources are are incomplete proteins complete proteins Beans are an exception to • Beans contain some this generalizations complete protein as well 3 Essential Amino Acids in Humans • Required in diet • Humans incapable of forming requisite carbon skeleton • Arginine* • Lysine • Histidine* • Methionine • Isoleucine • Threonine • Leucine • Phenylalanine • Valine • Tryptophan * Essential in children, not in adults 4 Non-Essential Amino Acids in Humans • Not required in diet • Can be formed from ααα-keto acids by transamination and subsequent reactions • Alanine • Glycine • Asparagine • Proline • Aspartate • Serine • Glutamate • Cysteine (from Met *) • Glutamine • Tyrosine (from Phe *) * Essential amino acids 5 Essential and Nonessential Amino Acids Nonessential Essentia l Alanine Arginine * Asparagine Histidine * Aspartate Isoleucine Cysteine Leucine Glutamate Lysine Glutamine Methionine Glycine Phenylalanine Proline Threonine Serine Tyrptophan Tyrosine Valine Amino acids are classified as glucogenic or ketogenic
    [Show full text]
  • Faculty of Engineering & Technology
    FACULTY OF ENGINEERING & TECHNOLOGY Dr. NIHARIKA SINGH Assistant Professor Dept. of Biotechnology Course: B. Sc Biotechnology Semester: 3rd Sub Code: CBBS-303 Sub Name: Biochemistry and Metabolism LECTURE 1 Dr. NIHARIKA SINGH Assistant Professor Dept. of Biotechnology INTRODUCTION Amino acids are a group of organic compounds containing two functional groups-amino and carboxyl. The amino group (-NH) is basic while the carboxyl group – (-COOH) is acidic in nature. General structure of amino acids: The amino acids are termed as α-amino acids, if both the carboxyl and amino groups are attached to the same carbon atom. The α-carbon atom binds to a side chain represented by R which is different for each of the 20 amino acids found in proteins. The amino acids mostly exist in the ionized form in the biological system. https://www.docsity.com/es/amimoacidos/3979454/ HISTORY The first amino acid which was discovered is Asparagine in 1806. Threonine was the last amino acid to be found in the year 1938. All the amino acids have trivial or common name from which they were first isolated. Asparagine was found in asparagus and glutamine was found in wheat gluten: tyrosine was first isolated from cheese and glycine (greek glykos means sweet) was so named because of the sweet taste. CLASSIFICATION OF AMINO ACID A. Nutritional classification of amino acid B. Classification of amino acid based on polarity C. Amino acid classification based on their metabolic fate D. Amino acid classification based on the structure E. Two main groups of amino acids 1. Essential or indispensable amino acid A.
    [Show full text]
  • The Emerging Role of Amino Acids of the Brain Microenvironment in the Process of Metastasis Formation
    cancers Review The Emerging Role of Amino Acids of the Brain Microenvironment in the Process of Metastasis Formation Francesca Cutruzzolà †, Amani Bouzidi †, Francesca Romana Liberati, Sharon Spizzichino, Giovanna Boumis, Alberto Macone , Serena Rinaldo , Giorgio Giardina and Alessio Paone * Laboratory Affiliated to Istituto Pasteur Italia, Department of Biochemical Sciences A. Rossi Fanelli, Sapienza University of Rome, Piazzale Aldo Moro 5, 00185 Rome, Italy; [email protected] (F.C.); [email protected] (A.B.); [email protected] (F.R.L.); [email protected] (S.S.); [email protected] (G.B.); [email protected] (A.M.); [email protected] (S.R.); [email protected] (G.G.) * Correspondence: [email protected]; Tel.: +39-0649910713 † These authors contributed equally to the work. Simple Summary: Why some cancers choose to form metastases in one organ rather than another is still largely unknown. In this review, we summarized the available information on the possible mechanisms controlling this choice. In particular, we tried to understand how some molecules, especially amino acids, released into the environment outside the cells, participate in selecting the brain as a target organ for the formation of metastases by specific types of aggressive tumors such as melanoma, breast, and lung cancer. Abstract: Brain metastases are the most severe clinical manifestation of aggressive tumors. Melanoma, Citation: Cutruzzolà, F.; Bouzidi, A.; breast, and lung cancers are the types that prefer the brain as a site of metastasis formation, even if the Liberati, F.R.; Spizzichino, S.; Boumis, reasons for this phenomenon still remain to be clarified. One of the main characteristics that makes a G.; Macone, A.; Rinaldo, S.; Giardina, cancer cell able to form metastases in the brain is the ability to interact with the endothelial cells of the G.; Paone, A.
    [Show full text]
  • Histidine: a Systematic Review on Metabolism and Physiological Effects in Human and Different Animal Species
    nutrients Review Histidine: A Systematic Review on Metabolism and Physiological Effects in Human and Different Animal Species Joanna Moro 1, Daniel Tomé 1, Philippe Schmidely 2, Tristan-Chalvon Demersay 3 and Dalila Azzout-Marniche 1,* 1 AgroParisTech, Université Paris-Saclay, INRAE, UMR PNCA, 75005 Paris, France; [email protected] (J.M.); [email protected] (D.T.) 2 AgroParisTech, Université Paris-Saclay, INRAE, UMR0791 Mosar, 75005 Paris, France; [email protected] 3 Ajinomoto Animal Nutrition Europe, 75017 Paris, France; [email protected] * Correspondence: [email protected]; Tel.: +33-1-44087244 Received: 1 April 2020; Accepted: 8 May 2020; Published: 14 May 2020 Abstract: Histidine is an essential amino acid (EAA) in mammals, fish, and poultry. We aim to give an overview of the metabolism and physiological effects of histidine in humans and different animal species through a systematic review following the guidelines of PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses). In humans, dietary histidine may be associated with factors that improve metabolic syndrome and has an effect on ion absorption. In rats, histidine supplementation increases food intake. It also provides neuroprotection at an early stage and could protect against epileptic seizures. In chickens, histidine is particularly important as a limiting factor for carnosine synthesis, which has strong anti-oxidant effects. In fish, dietary histidine may be one of the most important factors in preventing cataracts. In ruminants, histidine is a limiting factor for milk protein synthesis and could be the first limiting AA for growth. In excess, histidine supplementation can be responsible for eating and memory disorders in humans and can induce growth retardation and metabolic dysfunction in most species.
    [Show full text]