A genetic disease characterized by profound deafness and retinitis pigmentosa that leads to blindness, caused in some cases by a defective myosin protein.
A fluid-filled sac enclosed by a lipid-bilayer membrane. A property of a motor protein or other enzyme that undergoes many reaction cycles before dissociating from its track or substrate. False - Evolution tends to conserve the structures of proteins rather than their sequences. What are the two conformations of hemoglobin?
How does BPG decrease the affinity of hemoglobin for oxygen? BPG binding to hemoglobin stabilizes the deoxy conformation. Structurally, myoglobin and hemoglobin are very similar proteins. In which of the following levels of structure do they differ most? Quaternary structure. In its interaction with hemoglobin, oxygen is:.
Which of the following best describes the tertiary structure of myoglobin? BPG aids oxygen delivery to tissues by increasing the affinity of myoglobin for oxygen. Fetal hemoglobin has a higher affinity for oxygen than maternal hemoglobin.
Which of the following statements correctly outlines the mechanism behind this observation? Which of the following statements correctly describes the interaction between an allosteric protein and an allosteric effector? A mutation that significantly affects the folding or function of a protein cannot be considered conservative.
If a Lys residue that interacts with 2,3-bisphosphoglycerate BPG in the central cavity of hemoglobin is changed to a Ser residue, how would this affect hemoglobin behavior? The T state would be less stable. Which of the following triggers the transition from T state to R state low to high affinity in hemoglobin? Myoglobin contains all three types of secondary structure. A newly-identified protein has a sigmoidal curve in a graph of fractional saturation versus ligand concentration. What can be deduced about this protein?
The protein binds the ligand cooperatively. What type of allosteric effector is BPG? BPG is a heteroallosteric effector because it alters the ability of hemoglobin to bind another ligand, oxygen. Term A single operon. Definition usually contains all the enzymes which are specific for synthesis of a special biomolecule.
Term A plasmid. Definition A small circular DNA that is not part of a bacterial chromesome. Term Which inhibitor of protein synthesis competes with aminoacyl-tRNAs for binding to the A-site of the ribosome?
Definition puromycin. Term Linear eukaryotic DNA molecules. Definition have many origins of synthesis while circular prokaryotic DNA usually have only one. Term The majority of protein synthesis occurs in the.
Definition cytoplasm. Term All are stages in transcription except. Term All are components required for peptide chain initiation except. Definition a Shine-Delgarno Sequence. Term Response elements. Definition are enhancers of transcription activated by metabolic factors. Term The DNA double helix is stabilized by. Definition both hydrogen bonds between bases base-pairing and hydrophobic and van der waals between stacked bases. Term The typical composition of a ribosome is about. Term The replication of DNA to form daugther strands is.
Definition semiconservative. Definition proofreading. We have known for some time that the number of SDA families of a given size does not follow a linear power law, whereas the number of MDAs does 39 Fig. S2 shows that new sequence profiles defined in the last year characterize sequences deposited in the NR database decades ago. Although the fraction of MDA families with a particular number of members has a power-law dependence on the family size as shown by the linear log—log plotsthe fraction of SDA families with a particular number of members does not.
I provide two illustrations of the protein universe Fig. Illustrations of sequence space in which area is proportional to the number of sequences or sequence families in that region. Top Left The unique sequence universe contains all sequence families.
Thirty-two percent of SDA sequence families have a known structure, with one-fifth of these from structural genomics. Top Right The repetitious sequence universe contains all sequences. Middle Numbers of sequences in the corresponding regions of Top Right. Bottom Numbers of families in the corresponding regions of Top Left. The number of different MDA families, which are different combinations of SDAs, can clearly expand with the number of sequences. The slow growth of SDAs and the leveling off seen in Fig.
Given the limited sensitivity of methods to recognize homology in sequences, those already at the limits of detection easily can be imagined to drift apart further to give rise to a new SDA family not recognized by an existing sequence profile. Leave a Comment Cancel reply. Proline is incorporated during protein synthesis; posttranslational processing adds the hydroxyl group. The isoleucyl-tRNA synthetase has a proofreading function that improves the fidelity of the aminoacylation reaction, whereas the histidyl-tRNA synthetase lacks such a proofreading function.
Explain why. Isoleucyl-tRNA synthetase sometimes catalyzes the addition of valine to tRNA Ile , an amino acid that is similar to but smaller than isoleucine and can readily fit into the synthetase active site. Histidine has no close structural analogs among the amino acids, greatly lowering the chance that tRNA His will be charged with the incorrect amino acid. In each case, the mutation allows proper folding of the protein and binding of GTP, but does not allow GTP hydrolysis.
At what stage would translation be blocked by each mutant protein? IF the 70S ribosome would form, but initiation factors would not be released and elongation could not start. Some aminoacyl-tRNA synthetases do not recognize and bind the anticodon of their cognate tRNAs; they use other structural features of the tRNAs to impart binding specificity. The gene for a eukaryotic polypeptide of amino acid residues is altered so that the polypeptide has an N-terminal signal sequence recognized by SRP and an internal nuclear localization signal, beginning at residue Where is the protein likely to be found in the cell?
In addition, it has been found that the improved Nanobody of SEQ ID NO: l and its variants are particularly suited for extending the half-life of immunoglobulin single variable domains that contain more than one disulphide bridge, such as VHH's and Nanobodies belonging to the "VHH-1 class" which as further described herein may comprise two or even three disulphide bridges. Again, this will become clear from the further description and the Experimental Part herein.
These and other advantages, as well as the various aspects, embodiments, uses and applications of the invention, will become clear from the further description herein. Reference is for example made to EP 0 and EP 0 , as well as the various published patent applications from Ablynx N. Ablyn N. It is also generally known all VHH's and Nanobodies contain at least one disuiphide bridge, between the cysteine residue at position 22 and the cysteine residue at position 92 numbering according to Kabat, see the patent applications of Ablynx N.
Although most VHH's contain only this single disul. Some non-limiting examples of such VHH-1 type Nanobodies given as illustration only: other VHH-1 type sequences against other targets can be found in some of the other patent applications from Ablynx N. For example and without limitation, whereas expression levels of more than 0. As mentioned, in a first aspect, the invention relates to an amino acid sequence that is directed against human serum albumin, which essentially consists of or is the following amino acid sequence:.
This amino acid sequence is also referred to herein as "Alb" or the "amino acid sequence of the invention". The amino acid sequence of Alb- 1 is:. In the above sequence, the main differences between Albl and Alb have been indicated in bold and underlined.
Thus, in a further aspect, the invention relates to an amino acid sequence that is a variant of the sequence Alb- 1 SEQ ID NO:2 , which variant comprises:.
As will be clear to the skilled person, and although the use of Alb is generally preferred within the context of the present invention, the variants described in the preceding paragraphs share with Alb some of the same amino acid substitutions i.
For these reasons, these variants are also referred to herein as "Alb! In one specific, but non-limiting aspect, an Alb variant is such that, when it is used in the storage stability assay described in Example 5 i. The variants of SEQ ID NO's 6 to 1 1 or other Alb variants with 1 to 3 amino acid residues at the C-terminus, which may each be independently chosen from naturally occurring amino acid residues and may for example be independently chosen from A, G, V, L and I may in particular be used when the albumin-binding Nanobody is provided at the G-terminal end of the polypeptide or protein construct.
Thus, in one specific, but non-limiting aspect, the invention provides proteins or polypeptides that essentially consist of the amino acid sequence Alb or of one of the Alb- 23 variants described herein. As further described herein, the amino acid sequence Alb and the further Alb variants described herein can be used with advantage as a moiety, binding unit or fusion partner in order to increase the half-life of therapeutic moieties such as polypeptides, proteins, compounds including, without limitation, small molecules or other therapeutic entities.
Thus, in another aspect, the invention provides polypeptides, proteins, constructs, compounds or other chemical entities that comprise or essentially consist of the amino acid sequence Alb or of one of the Alb variants described herein and one or more other amino acid sequences, binding domains, binding units or other moieties or chemical entities.
In particular, the invention provides polypeptides, proteins, constructs, compounds or other chemical entities that comprise the amino acid sequence Alb or of one of the Alb variants described herein and one or more such as one or two therapeutic moieties which. Such polypeptides, proteins or constructs may for example and without limitation be a fusion protein, as further described herein. The invention further relates to therapeutic uses of such polypeptides, proteins, constructs or compounds and to pharmaceutical compositions comprising such polypeptides, proteins, constructs or compounds.
In one aspect, the at least one therapeutic moiety comprises or essentially consists of a therapeutic protein, polypeptide, compound, factor or other entity. In another embodiment, the at least one therapeutic moiety comprises or essentially consists of a therapeutic protein or polypeptide.
Comb Chem High. In yet another aspect, the at least one therapeutic moiety comprises or essentially consists of an antibody variable domain, such as a heavy chain variable domain or a light chain variable domain.
In a preferred aspect, the at least one therapeutic moiety comprises or essentially consists of at least one immunoglobulin single variable domain, such as a domain antibody, single domain antibody, "dAb" or Nanobody such as a VHH, a humanized VHH or a camelized VH or an IgNAR domain.
In a specific embodiment, the at least one therapeutic moiety comprises or essentially consists of at least one monovalent Nanobody or a bivalent, multivalent, bi specific or muitispecific Nanobody construct. The polypeptides, fusion proteins, constructs or compounds that comprise Alb or an Alb variant and one or more therapeutic moieties will generally and preferably have an increased half-life, compared to the therapeutic moiety or moieties per se. Generally, the constructs or fusion proteins described herein preferably have a half-life that is at least 1.
Also, preferably, any such fusion protein or construct has a half-life in man that is increased with more than 1 hour, preferably more than 2 hours, more preferably of more than 6 hours, such as of more than 12 hours, compared to the half-life of the corresponding therapeutic moiety per se. Also, preferably, any fusion protein or construct has a half-life in man that is more than 1 hour, preferably more than 2 hours, more preferably of more than 6 hours, such as of more than 12 hours, and for example of about one day, two days, one week, two weeks or three weeks, and preferably no more than 2 months, although the latter may be less critical.
Methods for pharmacokinetic analysis and determination of half-life are familiar to those skilled in the art. As mentioned, in one aspect, the amino acid sequence Alb or of one of the Alb variants described herein can be used to increase the half-life of one or more. Thus, one embodiment of the invention relates to a polypeptide, construct or fusion protein that comprises the amino acid sequence Alb or one of the Alb variants described herein and one or more such as one or two immunoglobulin single variable domain sequences, which are suitably linked to each other, either directly or optionally via one or more suitable linkers or spacers.
As mentioned herein, each such immunoglobulin single variable domain present in such a polypeptide, construct or fusion protein may independently be a domain antibody, single domain antibody, "dAb' :!
Preferably, each such immunoglobulin single variable domain is a Nanobody; and according to one specific but non-limiting aspect, at least one and up to all of these immunoglobulin single variable domains is a Nanobody of the VHH-1 class.
For example and without limitation, such a construct, fusion protein or polypeptide may comprise:. Some non-limiting examples of constructs, fusion proteins or polypeptides of the invention can be schematically represented as follows, in which "[Alb]" represents Alb or of one of the Alb variants described herein , "[therapeutic moiety 1 ] "" and "[therapeutic moiety 2]" represent the therapeutic moieties which as mentioned may each independently be an immunoglobulin single variable domain , " - " represents a suitable linker which is optional and the N-terminus is on the left hand side and the C-terminus is on the right hand side:.
When two or more different therapeutic moieties such as two or more different immunoglobulin single variable domains are present in the constructs or polypeptides of the invention, they may be the same or different, and when they are different they may be directed towards the same target for example, to the same or different parts, domains, subunits or epitopes of said target or to different targets.
Again, Alb and the other Nanobodies may be suitably linked to each other either directly or optionally via one or more suitable linkers or spacers, and according to one specific but non-limiting aspect at least one and up to all of the other Nanobodies may be of the VHH- 1 class.
For a general description of multivalent and multispecific polypeptides containing one or more Nanobodies and their preparation, reference is also made to Conrath et ai, J, Biol. Thus, in one specific but non-limiting aspect, the invention relates to a polypeptide or protein construct that comprises or essentially consists of Alb preferred or an Alb variant as described herein , which is suitably linked either directly or via one or more suitable linkers to one or two Nanobodies against c-Met.
As mentioned, according to a specific but non-limiting aspect, said one or two Nanobodies against c-Met comprise two disulphide bridges i. Many of the epidemics erupt during the intermediate stages of seasons of cold and hot climatic temperatures, humid 5 and non-humid conditions etc, Even the fluctuating the moods of the mind also will influence the health.
Hence maintaining equilibrium in every state of life is essential. The-flexibility property of the human being will give tolerance-against thesevariations hence person who possess this property will be usually healthy and happy. Hence maintaining healthy levels of energy will lead to healthy condition for which 10 different molecules with energy absorbing, conditioning and donating properties will be useful.
The behavior of a molecule under different conditions like temperature, pH, viscosity, ionic nature of the media in which the molecule is present can be understood. In a set of animals, which are maintained under experimental conditions, may have some commonality in the response.
But practically in an un controlled conditions the same response cannot be 20 observed. Hence the medicine tested in controlled conditions may differ in the day-to day life of the humans in uncontrolled conditions.
The study of the response of the chemical and bio chemical reactions could be tested under practical conditions. In the animated figure the same is shown. The radiations when moved with respect to time the quantum of energy will not be the same. Similarly a molecule having a 25 particular quantum of energy will vary in its energy when it is exposed to different temperatures, pH and Ionic media and give different results from person to person and place to place, so on.
Even though the medicine is consumed at single time various constituents in it will be moving in different speeds due to their interaction with the surface on it is moving, like a set of molecules get separated over a chromatographic 30 surface. It is the final quantum of energy being able to be measured which actually brings a change in the chemical atmosphere. Thus measurement of the energy dealt by a molecule along with its electrical charge will help to understand the chemical and therapeutic property of the sample under test.
The fingerprints of medicines with a specific color were given. The relation of color with efficacy was mentioned in traditional medicines. The color of absorbance is due to the chemical constituents present in it. The transmitted color of the sample was used as an indicator for the efficacy of the medicine.
Thus indirectly the color of absorbance is 5 used for the said efficacy. The fingerprints of different medicines with a specific taste were given in different figures. The order of taste is foundto be the order of chemical constituents in a specific order of polarity. Hence taste classification of medicines is the classifications based on polarity of the chemical constituents. The medicines will possess the required 10 efficacy if they contain constituents having required polarity along electromagnetic radiation properties qualitatively and quantitatively.
The three Highly Bitter medicines were fingerprinted. Substitution of single medicines is common in commercial market assessment of right variety will help to select and used to achieve better clinical uses. In a state of unconformity fingerprints is will help to identify the better variety. Usually Swertia Chirata is substituted with Andrographis Paniculata.
It can be seen that the high polar constituents present in Swertia is not seen in Andrographis. Hence it cannot be used for Pitta hara properties. Thus the efficacy should be checked while substituting any medicine. The rich profile in the retention times of minutes with Bitter taste can be seen in all the samples. The medicines like Chitraka and Danti are mentioned to have a special property called "The Prabhava".
Even though the medicines contain all tastes the first is majorly Pitta Kaphahara and the second is Kapha Vatahara. So first will close the channels and the second open the channel. There are different types of Prabhava. The medicines like Rudraksha and Sahadevi were also told to be examples of Prabhava.
When the 25 Rudraksha was soaked for longer time more quantity of samples were found to be get extracted. Sahadevi is mentioned for the treatment of Cancer. Lekhaneeya medicines: When medicines used for a specific efficacy are analyzed and the fingerprints were studied the common molecules can be seen indicating efficacy. Charaka Dashaimani Jeevaneeya medicines: The fingerprints of medicines classified as Jeevaneeya Vitalizes were shown.
Molecules of specific polarity have been mentioned for a specific efficacy. Two generally used Medhya dravyas: fingerprints of Bacopa and Centella were presented. The Profile of Bacopa is more in Pitta and the profile in Centella is rich in 5 constituents. Different substitutions need to be standardized. When some of the Medhya Rasayana dravyas were observed a common chemical profile is-seen as show, marked. Thus different targeted efficacies were indicated in classifying the medicines based on efficacy rather than plat pharmacognostic properties.
Rasayana dravyas of Swasa Bronchial diseases Rasayana dravyas of Sthoulya Obesity Rasayana dravyas: Medicines like Gingokobiloba and Ashwagandha were considered as highly potent herbal Rasayana medicines. The similarity of two different plants for same efficacy will help for better substitutions. Rasayana dravyas in general found to have an array of constituents in the entire range of polarity. Hence commonly they will be wide acting medicines.
But medicines having molecules from are found to be the immunomodulators. Constituents from are anti oxidants. Finger prints of Different sources of Vidarigandha species: Different sources of Vidarigandha Ipomoea digitata shows variation of chemical assay of the constituents the common molecules present in all varieties show that all these have some 25 commonalities and variations. Finger prints of Different sources of Amra Gandhi Haridra species: Collection and Processing of medicines needs to be standardizes.
Herbal medicines collected from different soils, pealed and unpeeled show variations of chemical assay. Different sources of Akarakarabha were presented. This helps to identify different 30 types of the single medicine available in the world.
Some of the medicines are used for achieving a. The medicines presented are used in Indian Systems of medicine for having a male child. This process is called as Pumsavana in Ayurveda. The Jeemutha Lunar effect: The influence of lunar effect on the chemical constituents of plants was reported in traditional texts, one of such plants has been studied.
The plant is showing different molecules of different efficacy when collected during specific timing. This emphasizes the need of standardization while collecting 5 herbal medicines. If molecule similar to progesterone can be seen in the sample collected on the full moon day of a specific month. Fingerprints of Sea buck thorn:- Some of the herbal- material used- in day-to-day life will have many therapeutic properties. Standardization of such material; from different sources will help to select correct variety for clinical or nutritional purposes.
Fingerprints of different sources of Aegle marmalous fruit are presented. Usually the immature fruit is prescribed for clinical purposes.
The ripe fruit show toxic profiles. Thus the collection specifications need to be standardized. Fingerprints of Drynaria qurcifolia show a rich profile. It is used for Osteo Arthrites. In Tamil 'Mudu' means joint Vattukkal means Vata hara. Arthritis is due to 15 Vata, which will be cured by this medicine. Single medicines used for hepatitis: Some of the medicines used for hepatic disorders were shown; medicines having constituents at the required polarity are proved to be potent.
Fingerprints of some Indian leafy vegetables are shown. The leafy vegetables 20 have become rich sources of anti oxidants and immunomodulators. If they are a part of the life as food material the health is maintained well.
Genetically modified orange juice: When the foods and the medicines are modified by different methods they should not lose or change the properties as mentioned in traditional texts. If it happens the traditional philosophies of medicines will go erratic, 25 as they have been designed based on the properties of material having specific physicochemical properties.
The fingerprints of a genetically modified food product, the orange juices were presented in the figure. After genetic modification, if the products do not contain the same properties like the original with similar efficacy, the efficacy cannot be tested by traditional methods and so will act differently. If all herbal 30 medicines are genetically modified the traditional philosophies will go erratic leaving the countries in dilemma about the traditional medicines and foods being used in day to-day life.
Fingerprints of some anti stress medicines were presented which show common chemical constituents which possess common therapeutic properties. Fingerprints of unknown material: When some materials like Sodium cyanide was analyzed, the Physico- chemical properties of the material were studied using the 5 fingerprints as shown in the figure.
Each country can develop the native plants as their traditional medicine using the basic concepts of traditional medicine. As any herbal -medicine. Thus the method helps to confirm the presence of properties of a medicine whether it has all required properties to be a medicine, as mentioned in traditional texts.
Taste is one of the basic parameter used in traditional drug standardization. The order of taste is mentioned towards a specific efficacy of the material having the respective taste. If one can assess the taste of any material, which facilitates, understanding the 15 efficacy of it, the drug discovery becomes easy.
Taste being a subjective parameter, one needs a tool, which can give the taste of an unknown, unbiased. Taste even changes with person and his health. Tastes were related to polarity based on our method. The selection of a material of specific taste helps to select a material of specific polarity to deal with a specific disease, which is also related to polarity.
The Astringency 20 Kashaya and Pungent Katu are found to be to high polar, where the second is less polar to first one. The Madhura, in traditional terminology was mentioned as the post assimilated Vipaka condition of Sweet. Then it is Vata hara. The molecules at minutes indicate Pitta vridhi, very high polar molecules leading to hyper acidity this makes the rest of the molecules to get fast absorbed by the body.
The molecules around 30 minutes are indicating Bitter, Sour and Salty by taste. Being a salt it should be salty by taste.
High polar molecules seen in salts but not in all bitters confirm this. Or the salt or bitter 30 may be dominating each other. It was observed that the polarity difference of these bitter, salty and sour tastes is very narrow. Being an unpalatable toxic chemical it will be difficult to confirm by humans. It is not showing any sweet property as shown in the sweet example. The post-assimilated Vipaka status of this material was not studied due to many experimental limitations, but can be studied.
Many of the medicines, which are bitter, show similar molecules at the same retention time. The salts at very high concentrations 5 show sour taste. Thus the taste is related to the amount of energy, the molecules possess and the taste receptor it can trigger having a specific polarity.
So it is the quantum of energy it can deal with that plays role in the efficacy of the medicine, irrespective ofits structure, many times. So salts should be acting due to their crystalline structures of the atoms arranged in specific order and geometry, which makes them therapeutically 10 active. The polarity of the crystals could be controlled due to the geometrical arrangements of the ionic molecules in the crystal.
These crystalline molecules should be triggering the respective taste receptors, resulting to specific tastes. That is why a PDA detector was able to give spectra of salts also. This indicates the utility of the present invention for assessing the property of an unknown plant or material. Thus it 15 helps for assessment of the chemical and therapeutic unreported medicines.
Some of the medicines used for female fertility was presented.Aug 17, · Method: FFPE tonsil sections underwent manual dewaxing and antigen retrieval step. All subsequent steps of staining, antibody elution and imaging were automated on the microscope integrated microfluidic device. A single tissue section was stained sequentially for CD3, CD4, CD8, CD20, CD56, CD68, FOXP3, PD-1, PD-L1 and CK with mouse or rabbit primary antibody and corresponding Alexa .