Structural Replacement of Active Site Monovalent Cations by the ε-Amino Group of Lysine in the ATPase Fragment of Bovine Hsc70†,‡

Biochemistry ◽  
1998 ◽  
Vol 37 (20) ◽  
pp. 7456-7462 ◽  
Author(s):  
Sigurd M. Wilbanks ◽  
David B. McKay
Keyword(s):  
Active Site ◽  
Amino Group ◽  
Monovalent Cations

scholarly journals Bacterial pseudokinase catalyzes protein polyglutamylation to inhibit the SidE-family ubiquitin ligases

Science ◽  
2019 ◽  
Vol 364 (6442) ◽  
pp. 787-792 ◽  
Author(s):  
Miles H. Black ◽  
Adam Osinski ◽  
Marcin Gradowski ◽  
Kelly A. Servage ◽  
Krzysztof Pawłowski ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Host Cell ◽  
Crystal Structures ◽  
Active Site ◽  
Bond Formation ◽  
Ubiquitin Ligases ◽  
Carboxyl Group ◽  
Amino Group ◽  
Isopeptide Bond

Enzymes with a protein kinase fold transfer phosphate from adenosine 5′-triphosphate (ATP) to substrates in a process known as phosphorylation. Here, we show that the Legionella meta-effector SidJ adopts a protein kinase fold, yet unexpectedly catalyzes protein polyglutamylation. SidJ is activated by host-cell calmodulin to polyglutamylate the SidE family of ubiquitin (Ub) ligases. Crystal structures of the SidJ-calmodulin complex reveal a protein kinase fold that catalyzes ATP-dependent isopeptide bond formation between the amino group of free glutamate and the γ-carboxyl group of an active-site glutamate in SidE. We show that SidJ polyglutamylation of SidE, and the consequent inactivation of Ub ligase activity, is required for successful Legionella replication in a viable eukaryotic host cell.

scholarly journals Active-Site Monovalent Cations Revealed in a 1.55-Å-Resolution Hammerhead Ribozyme Structure

2013 ◽  
Vol 425 (20) ◽  
pp. 3790-3798 ◽  
Author(s):  
Michael Anderson ◽  
Eric P. Schultz ◽  
Monika Martick ◽  
William G. Scott
Keyword(s):  
Active Site ◽  
Hammerhead Ribozyme ◽  
Monovalent Cations

Nitrogen and Sulfur Transferases

2007 ◽  
Author(s):  
Perry A. Frey ◽  
Adrian D. Hegeman
Keyword(s):  
Steady State ◽  
Reaction Mechanism ◽  
Active Site ◽  
Reaction Mechanisms ◽  
Radical Reaction ◽  
Amino Group ◽  
Steady State Kinetics ◽  
Ping Pong ◽  
Polar Reaction

Unlike other group transfer reactions in biochemistry, the actions of nitrogen transferring enzymes do not follow a single unifying chemical principle. Nitrogen-transferring enzymes catalyze aminotransfer, amidotransfer, and amidinotransfer. An aminotransferase catalyzes the transfer of the NH2 group from a primary amine to a ketone or aldehyde. An amidotransferase catalyzes the transfer of the anide-NH2 group from glutamine to another group. These reactions proceed by polar reaction mechanisms. Aminomutases catalyze 1,2-intramolecular aminotransfer, in which an amino group is inserted into an adjacent C—H bond. The action of lysine 2,3-aminomutase, described in chapter 7, is an example of an aminomutase that functions by a radical reaction mechanism. Tyrosine 2,3-aminomutase also catalyzes the 2,3-amino migration, but it does so by a polar reaction mechanism. In this chapter, we consider NH2-transferring enzymes that function by polar reaction mechanisms. Transaminases or aminotransferases are the most extensively studied pyridoxal-5'-phosphate (PLP)–dependent enzymes, and many aminotransferases catalyze essential steps in catabolic and anabolic metabolism. In the classic transaminase reaction, aspartate aminotransferase (AAT) catalyzes the fully reversible reaction of L-aspartate with α-ketoglutarate according to fig. 13-1 to form oxaloacetate and L-glutamate. Like all aminotransferases, AAT is PLP dependent, and PLP functions in its classic role of providing a reactive carbonyl group to function in facilitating the cleavage of the α-H of aspartate and the departure of the α-amino group of aspartate for transfer to α-ketoglutarate (Snell, 1962). PLP in the holoenzyme functions in essence to stabilize the α-carbanions of L-aspartate or L-glutamate, the major biological role of PLP discussed in chapter 3. The functional groups of the enzyme catalyze steps in the mechanism, such as the 1,3-prototropic shift of the α-proton to C4' of pyridoxamine 5'-phosphate (PMP). The steady-state kinetics corresponds to the ping pong bi bi mechanism shown at the bottom of fig. 13-1. This mechanism allows L-aspartate to react with the internal aldimine, E=PLP in fig. 13-1, to produce an equivalent of oxaloacetate, with conversion of PLP to PMP at the active site (E.PMP), the free, covalently modified enzyme in the ping pong mechanism.

scholarly journals A convenient synthesis of labelled rhodopsin and studies on its active site

1970 ◽  
Vol 119 (3) ◽  
pp. 359-366 ◽  
Author(s):  
M. D. Hirtenstein ◽  
M. Akhtar
Keyword(s):  
Conformational Change ◽  
Sodium Borohydride ◽  
Active Site ◽  
Cetyltrimethylammonium Bromide ◽  
Chemical Reactivity ◽  
Amino Group ◽  
Polar Environment ◽  
Convenient Synthesis ◽  
First Time ◽  
Native Site

Digitonin solutions of labelled rhodopsin, containing 3H in the retinyl moiety, were prepared by two related methods. Labelled rhodopsin was also prepared for the first time in cetyltrimethylammonium bromide and purified by column chromatography. It was shown that only certain rhodopsin preparations on denaturation in the dark and the reduction with sodium borohydride gave up to 60% of the radioactivity in a fraction characterized as N-retinylphosphatidylethanolamine. Such preparations also gave a lipid-linked retinyl moiety at the metarhodopsin-I stage, but, as expected, a protein-linked retinyl moiety at the metarhodopsin-II stage. Other preparations however, gave exclusively protein-bound radioactivity at the native-rhodopsin, metarhodopsin-I and metarhodopsin-II stages. It is therefore conceivable that the formation of N-retinylphosphatidylethanolamine is due to a non-enzymic reaction resulting from the transfer of the retinyl moiety from its native site to an amino group of a favourably oriented phospholipid molecule. The only firmly established aspect of the rhodopsin active site remains the demonstration in our previous work that at the metarhodopsin-II stage the retinyl moiety is linked to an ∈-amino group of lysine. On the basis of chemical reactivity it is argued that the light-induced conversion of rhodopsin into metarhodopsin II involves a profound conformational change resulting in the dislocation of the retinylideneiminium chromophore from a non-polar environment in rhodopsin to a polar environment in metarhodopsin II.

scholarly journals The incorporation of tritiated retinyl moiety into the active-site lysine residue of bacteriorhodopsin

1979 ◽  
Vol 183 (1) ◽  
pp. 175-178 ◽  
Author(s):  
E Mullen ◽  
M G Gore ◽  
M Akhtar
Keyword(s):  
Active Site ◽  
Lysine Residue ◽  
Halobacterium Halobium ◽  
Amino Group ◽  
Purple Membranes

Purple membranes were isolated from Halobacterium halobium bleached and regenerated with all-trans-[15-3H]retinal. The incorporation of label was 1.2 mol of retinal/mol of bacterio-opsin. The [3H]retinyl-bacterio-opsin obtained from regeneration was hydrolysed to give tritiated retinyl-lysine, which, on hydrogenation to N-epsilon-perhydro[3H]retinyl-lysine and reaction with 1-fluoro-2,4-dinitrobenzene, gave bis-(2,4-dinitrophenyl)-N-epsilon-perhydro[3H]retinyl-lysine. This result confirmed that the retinyl moiety of the chromophore is attached to an epsilon-amino group of lysine.

pK of the lysine amino group at the active site of acetoacetate decarboxylase

Biochemistry ◽  
1971 ◽  
Vol 10 (7) ◽  
pp. 1249-1253 ◽  
Author(s):  
Frank H. Westheimer ◽  
Donald E. Schmidt
Keyword(s):  
Active Site ◽  
Amino Group ◽  
Acetoacetate Decarboxylase

scholarly journals Sortase C-Mediated Anchoring of BasI to the Cell Wall Envelope of Bacillus anthracis

2007 ◽  
Vol 189 (17) ◽  
pp. 6425-6436 ◽  
Author(s):  
Luciano A. Marraffini ◽  
Olaf Schneewind
Keyword(s):  
Cell Wall ◽  
Bacillus Anthracis ◽  
Active Site ◽  
Diaminopimelic Acid ◽  
Side Chain ◽  
Carboxyl Group ◽  
Amino Group ◽  
Active Site Cysteine ◽  
New Hosts ◽  
Host Death

ABSTRACT Vegetative forms of Bacillus anthracis replicate in tissues of an infected host and precipitate lethal anthrax disease. Upon host death, bacilli form dormant spores that contaminate the environment, thereby gaining entry into new hosts where spores germinate and once again replicate as vegetative forms. We show here that sortase C, an enzyme that is required for the formation of infectious spores, anchors BasI polypeptide to the envelope of predivisional sporulating bacilli. BasI anchoring to the cell wall requires the active site cysteine of sortase C and an LPNTA motif sorting signal at the C-terminal end of the BasI precursor. The LPNTA motif of BasI is cleaved between the threonine (T) and the alanine (A) residue; the C-terminal carboxyl group of threonine is subsequently amide linked to the side chain amino group of diaminopimelic acid within the wall peptides of B. anthracis peptidoglycan.

scholarly journals Determination of protonation states of iminosugar–enzyme complexes using photoinduced electron transfer

2017 ◽  
Vol 8 (11) ◽  
pp. 7383-7393 ◽  
Author(s):  
Bo Wang ◽  
Jacob Ingemar Olsen ◽  
Bo W. Laursen ◽  
Jens Christian Navarro Poulsen ◽  
Mikael Bols
Keyword(s):  
Electron Transfer ◽  
Active Site ◽  
Photoinduced Electron Transfer ◽  
White Rot ◽  
White Rot Fungus ◽  
Amino Group ◽  
Enzyme Complexes

N-alkylated analogues of 1-deoxynojirimycin inhibit β-glucosidase from white rot fungus. The amino group in the β-glucosidase–iminosugar complex is unprotonated when bound, while an active site carboxylate is protonated.

The reaction with nitrous acid of the "active site" N-terminal isoleucine in chymotrypsin and derivatives

1970 ◽  
Vol 48 (6) ◽  
pp. 671-681 ◽  
Author(s):  
Joan W. Dixon ◽  
T. Hofmann
Keyword(s):  
Active Site ◽  
Rate Constants ◽  
Nitrous Acid ◽  
Amino Group ◽  
Amino Groups ◽  
Guanidinium Chloride ◽  
Third Order ◽  
Potential Activity ◽  
Chymotrypsin B ◽  
Nitrophenyl Ester

Bovine α-chymotrypsin, δ-chymotrypsin, homoarginine-δ-chymotrypsin, and bovine chymotrypsin B were inactivated by nitrous acid at pH 3.8–4.4 and 0°. The potential activity of bovine chymotrypsinogen A was not affected under these conditions. The inactivation rates as measured with the substrates α-N-acetyl-L-tyrosine ethyl ester, carbobenzoxyglycine p-nitrophenyl ester, and p-nitrophenyl acetate, and by 3H-di-isopropyl phosphorofluoridate incorporation were identical with the deamination rates of the amino group of the N-terminal isoleucine-16, but were slower than the deamination rates of the amino groups of the N-terminals half-cystine-1 and alanine-149. It is concluded that the deamination of isoleucine-16 is directly responsible for the inactivation. Third-order deamination rate constants of the N-terminal isoleucine-16 were measured and the following values (in min−1M−2) were obtained: α-chymotrypsin, 0.4–0.6; homoarginine-δ-chymotrypsin, 0.05; di-isopropyl phosphoryl-α-chymotrypsin, [Formula: see text]; tosyl-α-chymotrypsin, 0.05; chymotrypsin B, 0.3; α-chymotrypsin in guanidinium chloride, 30–50; homoarginine-δ-chymotrypsin in guanidinium chloride, > 20. The deamination rate constants for the model dipeptides isoleucylvaline and valylvaline are 40 and 46, respectively (Kurosky, A., and Hofmann, T.: to be published). A comparison shows that the constants for the dipeptides and the two chymotrypsins in guanidinium chloride are very close and are probably those of a fully exposed amino group. The much lower constants for the other enzymes and derivatives represent the varying degrees of accessibility of the amino group and show the usefulness of the reagent as a conformational probe. The results are fully compatible with the proposed structure of α-chymotrypsin (1) and the proposed function of the N-terminal isoleucine (2).

Sign in / Sign up

Export Citation Format

Share Document