Purification of Alcohol Dehydrogenase From Bovine Liver

Purification of Alcohol Dehydrogenase From Bovine Liver Jekathjenani Ratnakumaran Namrata Verma Introduction: In the world of chemistry, there are millions of enzymes, but in this lab the enzyme used is bovine alcohol dehydrogenase. This enzyme occurs in various mammalian tissues, but generally found in high concentrations in the organs such as liver and kidney. According to its name Bovine alcohol dehydrogenase, which implicates that it is collectively formed from bovine (cow), alcohol and the enzyme dehydrogenase. The protein was extracted from the liver of bovine. Alcohol is an organic compound which contains carbon atom (single bonds) and hydrogen atoms. This alcohol is available in various forms of liquid and used for a variety of purposes. According to its properties, alcohol is a hydroxyl group which has a sweet odor similar to fruit. Alcohol are further divided and identified into different groups and also they are polar. As they possess hydrogen bonding they have higher boiling points. Dehydrogenase is a type of an enzyme which oxidizes a substrate by a reduction reaction that trans fers one or more hydrogen H- to the electron acceptor which is NAD+ /NADP (nicotinamide adenine dinucleotide) or FAD Flavin coenzyme (Shibusawa et al, 2004). Collectively, it forms alcohol dehydrogenase, which is ADH persuaded by ethanol and acetaldehyde as they relate to carbon catabolite repression. It is also zinc containing enzyme which is activated by glutathione and EDTA, which contains heavy metals (Pateman et al, 1983). Many organisms contain an alcohol dehydrogenase enzyme which catalyzes the NADPH dependent of aromatic and aliphatic aldehydes into subsequent alcohols and catalyze the reduction of glyceraldehyde to glycerol (Arslanian et al, 1971). Alcohol dehydrogenase contains a several isozymes which catalyze the oxidation of primary and secondary alcohols to convert into aldehydes and ketones (Arslanian et al, 1971). The molecular weight of this enzyme is 39677.13 Da and it is made up of 374 amino acid sequence. The monoisotopic mass of this enzyme is 39651.32 and its p H value ranges between 8.6 to 9.0 with an extension coefficient of 12.6 and an isoelectric point at 5.4, its theoretical pI is 7.46.The alcohol dehydrogenase is also known for its battle against alcohol , its toxic molecules which negotiates with the nervous system so, the body organs which consist of high toxic of alcohol are liver and stomach which converts alcohol to acetaldehyde which is even more toxic substance and further it is conversted to acetate which is utilized by the cells present within our body (Goodsell et al, 2001). So overall, alcohol dehydrogenase converts potentially dangerous molecule into food ustilized by the cells preent within the body.In human bidy, alcohol dehydrogenase can create upto nine different kinds of alcohol dehydrogenase each having different properties. For example in liver beta3 enzyme(Goodsell et al, 2001).These each enzyme is formed of two subunits and they can be mixed and match to create mixed dimerss which are more active.Alcohol dehdroge nates also modifies certain other alcohols with giving outcome of dangerous products such as methanol. These by products are converted into formaldehyde by help of alcohol dehydrogenase (Goodsell et al, 2001). Catalytic activity of alcohol dehydrogenase: NADPH + an aldehyde NADP+ + an alcohol Methods: In order to conduct this laboratory experiment, All the required apparatus and materials were provided during the lab. Certain precautions and safety rules were followed such as gloves, safety glasses and lab coat. This lab was conducted for about duration of 11-12 weeks. According to the article (Arslanian et al, 1971) most of the steps and procedure was followed. Purification of the enzyme was carried out by following up eight steps. Precautions were made while the experiment was performed. Equipments were rinsed with distilled water before starting the experiment. The reagent and buffer solution were prepared with distilled water. All procedures were carried out at 0-40C. Buffer Preparation: The first step involved in the buffer preparation. The stock solution, 0.1M Tris HCl at pH 7.6 was prepared by dissolving 12.14 g of Tris base with 1000 ml of distilled water and the pH was adjusted to 7.6 by adding diluted HCl. The Tris HCl buffer solution with different concentrations such as 10mM (pH 7.5), 40mM (pH 7.6) and 50mM (pH 7.5) were prepared by diluting the stock solution with distilled water. Sodium Chloride elutant buffer (0.16M NaCl) was prepared by dissolving 9.3504 g of NaCl with 1000 ml of distilled water. Preparation of Homogenate: The second step involved the preparation of the homogenate. The bovine liver was homogenized in a Waring blender in 90 ml of 0.32 M sucrose in 10mM Tris HCl buffer at pH 7.5. Approximately 27.39 g of sucrose was added in 250 ml of 0.01M Tris HCl to make 0.32M sucrose in 10mM of Tris HCl at pH 7.5. The homogenate was centrifuged at 15000 RPM for 30 minutes using centrifuge-Sorvall RC5 refrigerated centrifuge SS 34. Ammonium Sulfate Fractionation: Step three involved ammonium sulfate fractionation. The homogenate was 35% saturated and equilibrated with ammonium sulfate by dissolving 20.9 g of ammonium sulfate in 250 ml of distilled water. The supernatant was centrifuged at 15000 RPM for 30 minutes and the precipitate was discarded. Then, concentration was increased to 60% saturated ammonium sulfate by dissolving 16.4 g in 500 ml of distilled water. The suspension was centrifuged at 15000 RPM for 30 minutes. The obtained gray pellet was dissolved in 40mM Tris HCl buffer at pH 7.6. Then, the solution was dialyzed against 2L of 0.04M Tris HCl at pH 7.6 for 24 hours and again dialyzed with same buffer for another 24 hours. Performing DEAE-Sepharose Chromatography: The fourth step involved DEAE-Sephrose Chromatography. DEAE-Sepharose column that can hold up to 10ml volume was applied. The column was equilibrated by applying four times of 10ml of 40mM of Tris HCl, pH 7.6 buffer. About 10ml volume of centrifuged and dialyzed material was applied through the column. The column was washed with the same buffer (40mM of Tris HCl, pH 7.6) and then eluted by 40mM of Tris HCl with 50mM of NaCl. About 1 ml volume of twenty fractions of enzyme solution was collected using microfuge tubes. Enzyme Activity Assay: Fifth step involved measuring enzyme activity using a spectrometer. The enzymatic activity was initiated with 1mL of volume of blank solution containing 20  µl of distilled water, 10  µl 33mM of ethanol, 10  µl of 0.26mM of NAD+ and 960  µl of 0.1M of glycine buffer. Enzymatic assay activity was measured by taking total volume of 1000  µl containing 20 µl of enzyme solution, 10  µl 33mM of ethanol, 10  µl of 0.26mM of NAD+ and 960  µl of 0.1M of glycine buffer. The wavelength was set up at 340nm and measured using Cary 50s and 60s spectrometer. One unit of activity equals 1 µmol NADH produced per min based on the absorption coefficient of 6220 mol/l/cm for NADH at 340 nm. The above procedure was repeated for kinetic analysis and the range of ethanol concentration used was 20 to 25 mM. The observed data were fitted using Lineweaver -Burk kinetic plots. Gel filtration: The sixth step involved gel filtration. The enzyme was precipitated by 62% saturated ammonium sulfate and dissolve in 10ml of 50 M Tris-HCl, pH 7.5. The suspension was centrifuged for 20 min at 10000 RPM. Then, the column of Sephadex G-50 was run with 10 ml of enzyme solution. The column was equilibrated and washed with 50 M Tris- HCl buffer, pH 7.5. Then, the column was eluted by 50mM of Tris HCl with 50mM of NaCl. Around 10 fractions were collected at the rate of 1 ml/min in a microfuge tube. The highest highest specific activity fractions were precipitated by 62% saturation with ammonium sulfate. In the final step, the enzyme was redissolved in 5 ml of same buffer and apply to the column of Sephadex G-50 under the same conditions. Again, the highest specific activity fractions were precipitated by 62% saturation with ammonium sulfate. Performing CM-Sephadex chromatography: The precipitated enzyme was dissolved in 1 ml of potassium phosphate buffer contain 0.02M of NaCl, pH 7.0 and the enzyme solution was dialyzed against the same buffer for 2 hours. 10ml of non diffusible material was applied to a CM-Sephadex column. Then, the column was equilibrated with the same buffer. Then, the enzyme was eluted from the column with two column volumes of 0.16 M of NaCl (20ml). 10 fractions were collected and precipitated with 62% saturated ammonium sulfate. Bradford Assay: The eighth step involved Bradford Assay. The data (absorbance) observed from Bradford Assay Standards was used for calculating the mass of BSA in  µg. The final step used in this experiment was SDS PAGE method. About 20 µl of enzyme with loading buffer was loaded on the gel and by observing the gel, the mass of the protein was calculated. Results: Bovine Alcohol Dehydrogenase (ADH) was purified by following up few methods. The experimental results were observed and recorded for appropriate methods. Using DEAE Sepharose Chromatography, fractions were collected and all the fractions were appeared colorless. The enzymatic assay activity was measured at 340nm using spectrometer. The figure 1 indicates that the enzyme activity was increased by absorbing the NADH. The highest specific activity was selected based on the graph obtained in the enzyme kinetic activity. However, this method failed, resulting no increased activity. The enzyme kinetic activity had done for all the fractions, but none of them shown the accurate result. The graph obtained from the spectrometer does not show the increased activity of the enzyme to conclude the presence of protein. The result of the enzyme activities of collecting fractions was shown in figure 5, 6, 7, 8 and 9 respectively. However, the Bradford assay method was performed and the absorbance of the standards and the enzyme were recorded in the following tables. Based on these values, the graph of standard curve of absorbance versus mass of BSA was plotted. Table 1: The following table represents the recorded values of absorbance at 540 nm and calculated the mass of the BSA using the Bradford assay method Figure 1: Represents the enzymatic activity obtained from the purified protein ADH after ammonium fractionation method had performed and the peaks are showing that activity is increased. The above plot was obtained at 340 nm using Cary 50s-60s spectrometer and it was run as two parts for 4 minutes. Figure 2: Represents the standard curve of A595 versus mass of ADH protein obtained from the Bradford assay method. From the slope value obtained from the curve, the mass of the ADH protein was calculated. The mass of the protein calculated from the figure 2 is 4.766  µg and concentration of the protein is 4.766  µg / 25  µl SDS-PAGE method: Table 2: Represents the recorded values of SDS-PAGE method for the determination of molecular weight of the ADH purified protein Protein Molecular weight (Dalton) Log (Molecular Weight) Mobility (cm) Strand 1 60000 4.778 4.8 Strand 2 50000 4.699 5.1 Strand 3 40000 4.602 6.5 Strand 4 25000 4.398 7.0 Strand 5 20000 4.301 9.3 Figure 3: Represents the SDS-PAGE analysis of purified protein bovine Alcohol Dehydrogenase (ADH). The graph was plotted with log of molecular weight versus mobility of protein based on the SDS-PAGE values. Figure 4: Represents the single band on an SDS-PAGE gel (9th lane). This figure shown the proof of the protein ADH present in the enzyme solution and mass of the protein was calculated based on the obtained SDS-PAGE results. From the figure 3 and 4, the mass of the protein calculated is 345143.74 Da Figure 5: Represents the enzymatic activity obtained in the fraction 9th of the purified protein ADH and the peaks are obtained at 340 nm using Cary 50s-60s spectrometer. Figure 6: Represents the enzymatic activity obtained in the fraction 10th of the purified protein ADH and the peaks are obtained at 340 nm using Cary 50s-60s spectrometer. Figure 7: Represents the enzymatic activity obtained in the fraction 12th of the purified protein ADH and the peaks are obtained at 340 nm using Cary 50s-60s spectrometer. Figure 8: Represents the enzymatic activity obtained in the fraction 13th of the purified protein ADH and the peaks are obtained at 340 nm using Cary 50s-60s spectrometer. Figure 9: Represents the enzymatic activity obtained in the fraction 14th of the purified protein ADH and the peaks are obtained at 340 nm using Cary 50s-60s spectrometer. Discussion: According to the experimental study, the outcome results were not satisfying, so overall the experiment was not successful it failed. Based on the SDS-PAGE, ADH purified protein was not much visible clearly on the gel. Proteins are viewed as bands. SDS-PAGE results indicates that smaller protein molecules are at the bottom of the gel and larger molecules are at the top of the gel. This is showing that SDS-PAGE gel separate the protein molecules based on the size and mass of the protein. Most of the protein bands are viewed in between the molecular weight, 100 kDa and 30 kDa. Determining mass and purifying the protein, Bovine Alcohol Dehydrogenase using the Bradford assay and SDS-PAGE procedure was conducted successfully using this experiment. The result obtained in the SDS-PAGE and Bradford Assay are differ from the standard value and the concentration of the protein was determined using these methods. Based on the molecular mass on the EXPASY website, the standard molecular mass of the ADH protein is 39677.13 Da. The experimental mass of the ADH protein is 345143.74 Da. The mass difference is a large number. This could occur due to the experimental errors. The experimental errors can be avoided by handling equipments and following the instructions in a proper manner. Predicting the protein band on SDS-PAGE gel could cause the error. Moreover, the purification method such as DEAE Sepharose Chromatography was performed to test the enzyme activity of the protein. The obtained results are shown as a figure 5, 6, 7, 8 and 9 in the results section. The overall results obtained in these figures indicated that the experiment was not turned successful. The figure 5, 6, 7, 8 and 9 shown that enzyme activities are decreasing and wiggling. They are not constantly increasing or decreasing. Therefore, it was concluded that the purification of the enzyme was not turned positive and it could be due to the human errors occurred while conducting the experiment. This could be po ssible due to various reasons such as, during measurements for making the solution at the very beginning may be the concentration required was not appropriate, due to human error it was not properly mixed. It could also be possible that while grinding the liver , certain chunks of the liver were still not properly collected due to which the amount of liver used was not effective to obtain supportive and positive results.The variability of the results presented here is loss of certain atoms during the process of purification as their was no enzyme activity observed.The substrate studies of the alcohol dehydrogenase isolated from the bovine liver have demonstrated the hydrophobic site for binding alcohol (Arslanian et al, 1971). The article mentioned that the buffer that has a low ionic strength is used for the enzyme adsorbtion which caused the incomplete deactivation of enzymes and it was proven evidently (Arslanian et al, 1971). Moreover, as there is no enzyme activity measured in step 5 (DEAE- Sephrose Chromatography), the gel filtration and CM-Sephadex Chromatography method was not performed for this study. The enzyme purification might get succeeded if the study has performed these two methods. The article mentioned that gel filtrations on Sephadex G-100 has successive ability to separate the enzyme from non enzyme protein (Arslanian et al, 1971.) For further studies, more information is required before conducting the study as well as the time allotted was less, due to which it could suggest certain results and test were not done at the appropriate time. In conclusion, the study was conducted by following the method listed in the article. This studys report discussed the properties and successful method for the purification of enzyme, Bovine Alcohol Dehydrogenase. Even though article procedures were followed, errors occurred which resulted in deviations in results. However, the methods of Gel filtration and CM Sephadex Chromatography where successive but could not be conducted in this lab because the enzyme activity was limited after DEAE Chromatography was performed. More caution should have gained while conducting the experiment. It is emphasized that further research on enzyme purification method could improve the results and find success in the study. Appendix: Sample calculation 1: Volume of one microliter= 0.001mL Volume of 20 microliter= (0.001 ml x 20  µL) / 1  µL = 0.02 ml Therefore, mass of protein in 1mL of stock solution= 0.10 mg Mass of protein in 0.02 ml of stock solution = (0.10 mg x 0.02 ml) / 1 ml = 2 x 10-3 mg To convert mg to  µL, multiply by 1000, Mass of protein= 2 x 10-3 x 1000 = 2  µg Absorbance of the ADH purified Protein, y = 0.2544 Slope Line of equation: Y=mx+b Y= 0.0505 x + 0.0137 0.2544 = 0.0505 x + 0.0137 The mass of the protein, x = (0.2544 0.0137) /0.0505  µg = 4.766  µg Concentration of the protein, C = mass/ volume = 4.766  µg / 25  µl = 0.19  µg/  µl Total mass that recovered= Conc. X Total volume = 0.19 x 1000  µl = 190.64  µg SDS- PAGE method: Absorbance of the ADH purified Protein, y = 0.2544 Slope Line of equation: Y= mx+b Y= 6.0902 x + 33.982 0.2544= 6.0902 x + 33.982 X= (0.2544 33.982) / 6.0902 = 5.538 The mass of the protein = 105.538 = 345143.74 Da References: Arslanian,M.J., Pascoe,E,. and Reinhold,J.G., (1971) Rat Liver Alcohol Dehydrogenase.Dept. of Biochem.School of Medicine, American University of Beirut.125,1039-1047. Alcohol Dehydrogenase(ADH)The university of Minnesota Biocatalysis/Biodegradation Database.Calzyme. Lab.inc. Shibusawa,Y.,Fujiwara,T.,Shindo,H., andIto,Y. (2004) Purification of alcohol dehydrogenase from bovine liver crude extract by dye-ligand affinity counter-current chromatography, J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci.799(2):239-44. Pateman,J.A., Doy,C.H.,Olsen,J.E.,Norris,U., Creaser. E.H., and Hynes,M.(1983) Regulation of Alcohol Dehydrogenase (ADH) and Aldehyde Dehydrogenase (ALDDH) in Aspergillus nidulans.Proceedings of the Royal society.Bio.Sci.217, 243-264. Ward,W.W., and Swiatek,G.,(2009) Protein purification.The state University,, Scool of Environmental and Biology Science,Department of Biochem. And Microbio.76,1- 21. Goodsell,D.(2001) Alcohol Dehydrogenase.Molecule if the month. RCSB.Protein Data Bank.doi: 10.2210.

History Of Communication Essay -- essays research papers

  Ã‚  Ã‚  Ã‚  Ã‚     Ã‚  Ã‚  Ã‚  Ã‚  Since the beginning of time, people have had the need to communicate with one and other. The most common type of communication is speech, but you could not talk to someone who lived 20 miles away. Then written language was developed, people marked symbols on paper, stone, or whatever was available. Then hundreds of years passed, and people who wanted to share their ideas with people had to do allot of writing, until someone thought to make a writing machine. This machine is called the printing press.   Ã‚  Ã‚  Ã‚  Ã‚  Gutenberg's invention of the printing press is widely thought of as the origin of mass communication-- it marked Western culture's first viable method of disseminating ideas and infomation from a single source to a large and far-ranging audience. The story of print is a long and complax one. It may be too much to claim that print was the single cause of the massive social, political and psychological changes it is associated with. However, print did wield enormous influence on every aspect of European culture. Some historians suggest that print was instrumental in bringing about all the major shifts in science, religion, politics and the modes of thought that are commonly associated with modern Western culture.   Ã‚  Ã‚  Ã‚  Ã‚  Gutenberg foresaw enormous profit-making potential for a printing press that used movable metal type. Despite their rapid growth in numbers, secular scribes simply could not keep up with the commercial demand for books. Gutenberg also saw strong maket potential in selling indulgences, the slips of paper offering written dispensation from sin that the Church sold to fund crusades, new buildings and other projects devoted to expanding its dominance. In fact, press runs of 200,000 indulgences at a time were common soon after the handwritten versions became obsolete.   Ã‚  Ã‚  Ã‚  Ã‚  There were many different innovations since the first hand operated printing press. The Stanhope press, which was widely used for many years, still used a hand-operated screw to press print and paper, but it could print up to 250 sheets an hour. A considerable improvement was the Colombian press. In this press, the typical screw method was eliminated, and replaced with powerful hand levers.   Ã‚  &nbs... ...the negative side, wars are waged more easily, the scope of human conflict has been extended along telephone lines, the multi-generational household has been broken-up as living alone is no longer an experiment in isolation, and the time-space continuum seems to be compressed faster than previously thought possible (Brooks, 1976). On the other hand, the invention of the telephone has resulted in the rapid and diffuse dissemination of technical and scientific information, saved lives through links to emergency services, made possible the modern city through telephonic connections, increased the speed and ease with which information changes place, and accelerated the rate of scientific and technological change and growth in industry (Brooks, 1976).   Ã‚  Ã‚  Ã‚  Ã‚  Since the invention of the printing press, communication over distances has become much more feasable. The invention of the the telephone, computer, and the internet has made such an impact on our society. Now we are able to view tremendous multitudes of information from our own living room. The history of modern communication is still ongoing, and will continue to progress far into the future.

Radio Music Essay

Music, I can say, has played an important role to the lives of every human. It has been a source of inspiration and leisure on past times. When you celebrate, music never gets lost and when you need to be calmed, music is also relaxing. As the years pass, along with it are the changes in trend. Characteristics of radio music by then are different of the radio music we have today. Several genres of music flourished from 1950s to 1960s. The birth of Rock and Roll is also dated back significantly in the 1950s. Right now, there is a question asked whether or not people would still listen to the radio despite of the latest gadgets for music are on hand. Listening to the radios are threatened by the latest technology including iPods and mp3 players. Aside from the radio, music is also presented by music channels in the television where music videos are accompanying the song. Radio is the major source of entertainment for the Americans having almost every household own at least one radio. 93% of the Americans owned a radio during that time. It was in the year 1954 that a rock and roll song landed the hit charts. Rock and Roll was a product of several genres including indigenous American jazz, rhythm and blues, swing, and the gospel style. Its roots trace back to the black culture of that moment (Faculty). Another music that was played on the radios is called â€Å"covers† (Faculty). Covering Family Name 2 takes place when a person or a group records again a song that was already recorded by another individual or group. This term’s roots dates back in the year 1966 (Online Etymology Dictionary). Songs back then that were originally sung by black artists were being asked to be covered by white artists. There were a lot of advantages that a â€Å"cover† has. First, they can outsell the original version due to having greater resources and second, the quality of the sound is much better than that of the original version. Rock and Roll had more specific genres which include Rock-a-Billy, Doo Wop, and the â€Å"West Coast† sound. â€Å"Rockabilly† was a mixture of rhythm and blues and country music. The â€Å"Doo Wop sound† traces its origins in the urban North. It has been fondly called and described as 1950s’ â€Å"street corner† music. Under Doo Wop are the characteristics of using â€Å"nonsense syllables† as background rhythm, having emphasis on its harmony, and a different range of voices or parts. Came 1960s and a new kind of rock and roll emerged called â€Å"California Sound. † California is the place where it first started. Rock and roll’s decline happened in the later part of 1950s since companies started to promote â€Å"teen idols. † The themes of their songs are mostly about love (Faculty). Some of the famous and promising artists during this time are the following: Dean Martin, Patti Page, Chuck Berry, Little Richard, Eddie Cochran, Carl Perkins, Elvis Presley- Don’t Be Cruel, Jailhouse Rock and many others, Arthur â€Å"Big Boy† Crudup, â€Å"Big Mama† Thornton, Hank Williams, Sr. , Buddy Holly and the Crickets, Every Brothers- Bye Bye Love, Roy Orbison, Ricky Nelson, The Marcels- Blue Moon, The Diamonds, Little Eva- Locomotion, The Shirelles- Will You Still Love Me Tomorrow? , Neil Sedaka- Breaking Up is Hard to Do, and Paul Anka with his song Diana (Faculty). Family Name 3 At the present, since whatever the trend in America is being adopted by other countries, there are 17 formats that are now playing and being listened to. Ranking from the first, the following are Country, News/ talk/ information, Adult Contemporary, Pop Contemporary Hit Radio, Classic Rock, Rhythmic Contemporary Hit Radio, Urban Contemporary, Urban Adult Contemporary, Oldies, Hot Adult Contemporary, Mexican Regional, Contemporary Christian, All Sports, Alternative, Classic Hits, Classical, and Talk/ Personality (Radio Today). From Rock and Roll, it can be seen from the above information that the taste of music listeners have shifted to many other kinds. In my own observation, today’s folks are getting more acquainted with music that have lively danceable beats but not necessarily with high notes. A lot of the radio stations would play Pop, R&B and Hiphop. The young is the targeted market of these radio stations. Leading artists of today include Beyonce Knowles, Neyo, Chris Brown, Rihanna, Akon, Taylor Swift and many others who have hit the charts just like those mentioned artists in the year 1950s and 1960s. Works Cited â€Å"Cover. † Online Etymology Dictionary. 5 May 2009. < http://www. etymonline. com/index. php? l=c&p=28/> â€Å"The Rise of Rock ‘n Roll. † Faculty. 5 May 2009. < http://faculty. smu. edu/dsimon/Change-Music. html/> â€Å"Radio Today. † Arbitron. 5 May 2009. < http://www. arbitron. com/downloads/radiotoday07. pdf/>

Summary of The Great Gatsby

Published in 1925, F. Scott Fitzgeralds The Great Gatsby is frequently studied in American literature classrooms (college and high school). Fitzgerald used many of the events from his early life in this semi-autobiographical novel. Hed already become financially successful with the publication of This Side of Paradise in 1920. The book is listed on the Modern Librarys list of 100 Best Novels of the 20th Century. Publisher Arthur Misener wrote: I think it (The Great Gatsby) is incomparably the best piece of work you have done. Of course, he also said that the novel was somewhat trivial, that it reduces itself, in the end, to a son of anecdote. Some of the very elements that brought the book acclaim were also the source of criticism. But, it was (and still is) considered by many to be one of the great masterworks of the time period, and one of the great American novels. Description Title: The Great GatsbyAuthor: F. Scott FitzgeraldType of Work Genre: Modernist Novel; FictionTime Place (Setting): Long Island and New York City; Summer 1922Publisher: Charles Scribners SonsPublication Date: April 10, 1925Narrator: Nick CarrawayPoint of View: First and Third Person Basics Great American literary classicOne of F. Scott Fitzgeralds most famous worksChronicled 1920s America, the Jazz AgeChallenged at the Baptist College in Charleston, SC (1987): language and sexual referencesThe first novel that Scribners had published that contained foul language. How It Fits In The Great Gatsby is usually the novel for which F. Scott Fitzgerald is best remembered. With this and other works, Fitzgerald forged his place in American literature as the chronicler of the Jazz Age of the 1920s. Written in 1925, the novel is a snapshot of the time period. We experience the glittery-splendiferous world of the wealthy—with the accompanying emptiness of morally decayed hypocrisy. Gatsby represents so much that is seductive, but his pursuit of passion—at the expense of all else—leads him to his own ultimate destruction. Fitzgerald writes: I wanted to get out and walk eastward toward the park through the soft twilight, but each time I tried to go I became entangled in some wild, strident argument which pulled me back, as if with ropes, into my chair. Yet high over the city our line of yellow windows must have contributed their share of human secrecy to the casual watcher in the darkening streets... I saw him too, looking up and wondering. I was within and without. Do you ever feel within and without? What do you think it means? Characters Nick Carraway: A Midwesterner, who sells bonds. Narrator. He observes and describes the rise and fall of Jay Gatsby.Daisy Buchanan: Wealthy. Cousin of Nick Carraway. Tom Buchanans wife.Tom Buchanan: Wealthy. Philanderer. Daisy Buchanans husband. Powerful personality.Jay Gatsby: A self-made man. The epitome of American Dream. A fascinatingly unforgettable figure in American literature. His parents were poor farmers. After getting a taste for wealth, he went into the Army, attended Oxford and quickly accumulated wealth via nefarious means. With his stupendous rise to great fortune, he was fated to fall.Jordan Baker: Daisys friend.George Wilson: Myrtle Wilsons husband.Myrtle Wilson: Tom Buchanans mistress. George Wilsons wife.Meyer Wolfsheim: An underworldly, criminal figure. Jay Gatsbys acquaintance.