JASCO CD Spectrometer by SUNWAY SCIENTIFIC CO.

Circular Dichroism spectropolarimeter

determining whether a protein is folded, and if so characterizing its secondary structure, tertiary structure

comparing the structures of a protein obtained from different sources or comparing structures for different mutants of the same protein

studying the conformational stability of a protein under stress -- thermal stability, pH stability, and stability to denaturants

determining whether protein-protein interactions alter the conformation of protein.

http://www.ap-lab.com/circular_dichroism.htm#CD_secondary

What is protein?

Proteins are large organic compounds made of amino acids arranged in a linear chain and joined by peptide bonds.

Proteins are essential parts of all living organisms and participate in every process within cells.

(abstract from WIKIPEDIA)

Structure of Protein

Primary Structure : Amino acid sequence

Structure of Protein

Secondary Structure : Arrangement of chain about an axis

Alpha Helix Beta Sheet

Structure of Protein

Tertiary Structure : Spatial relationship of amino acid units that are relative far apart in the protein chain

Denaturation and aggregation of proteins

denaturation and aggregation of proteins is also a problem with serious medical implications, e.g. in human diseases like Alzheimer’s, Parkinson’s, etc.

protein denaturation during production, shipping, storage, and delivery of therapeutic proteins is a problem of significant economic importance.

(abstract from WIKIPEDIA)

Optical Rotary Dispersion

Optical Rotary Dispersion

ORD is the difference in refractive indices of the

medium for the two circular polarizations

Optical Rotary Dispersion

The plane of polarized light rotates by a

chiral molecule is called optical rotary

(Polarimeter).

The variation of optical rotation as a function

of wavelength is called optical rotary

dispersion (ORD).

Circular Dichroism

Circular Dichroism

Circular Dichroism

Circular Dichroism

Circular Dichroism

Circular Dichroism

Circular Dichroism

Circular Dichroism

Circular Dichroism

CD is the difference in absorption coefficients of an optical active medium for left and right circularly polarized light

Far UV CD Secondary Structure Estimation

Biochimica et Biophysica Acta 1751 (2005) 119 – 139

Near UV CD Tertiary Structure Determination

Biochimica et Biophysica Acta 1751 (2005) 119 – 139

Protein Identification

CD spectra of D-amphetamine (blue) and Dmethamphetamine(red).

Chemical denaturation

Myoglobin structure changes during an automated pH titration

Thermal denaturation

Thermal Denaturation of Hen-egg Lysozyme from 20 to 95°C.

Thermal denaturation

Analyzing the heating history of the insect

contaminant in a beverage container

JASCO Corporation was founded in 1958

Founding members included World

famous physicist Yoshio Fujioka and

Shinichiri Tomonaga, The Nobel Prize

winner.

Jasco through history

1961 AP-1 first JASCO Spectropolarimeter (ORD only)

1963 ORD/UV-5 ORD/UV capabilities in the same unit, widely

sold worldwide

1965 ORD/UV-5 with CD option, first CD in JASCO

1970 J-20 ORD/CD Spectropolarimeter

1972 J-40 first dedicated CD unit (at the beginning still

with Pockel cell)

first data processors for spectra averaging …..

1978 J-500 refinement of J-40 design, updated electronics

and data processor(DP-500), later on

interface for MS-DOS computers (IF-500II)

The history of Jasco CD spectrometers

1980 J-200 NIR CD unit (800-2000 nm)

1986 J-600 fully PC controlled CD spectropolarimeter

(MS-DOS soft)

1990 J-710/720 widely renovated unit with PC control

(MS-DOS and later on Windows),followed

by J-730 for NIR

1995 J-715/J-710 update, with refinements

1999 J-810 compact model

2006 J-815 latest successful model

The history of Jasco CD spectrometers

J-815 CD Spectrometer

The latest efforts in the JASCO

commitment to lead the field of CD

Compact benchtop design

Optimum performance

Multiple acquisition mode

Wide ranges of accessories

Windows XP compatible Spectra Manager

software for control and data analysis

Compact benchtop design

Dimension: 1115 (W) × 576 (D) ×410 (H) mm

No external power supply unit J-720 : Main unit : 1230 (W) × 580 (W) × 400 (H)

plus power supply & AVR units

Optimum performance

WL range : 163 ~ 900 nm (170 ~ 800 nm)

Highest sensitivity : ~ 185 nm, 0.030 mdeg (RMS noise) (0.05mdeg) 200 nm ~, 0.020 mdeg (> 0.035mdeg)

Extra low stray light : < 0.0003 mdeg (NA)

Fast scanning speed : max. 10000 nm/min (5000 nm/min)

High CD resolution : 0.0005 mdeg (NA)

Multiple acquisition mode

Wavelength scan : Continuous scan Step scan Auto-scan

Time scan : Fixed WL time scan

Temperature scan : Fixed WL temp. scan 3D scan (CD vs. WL vs. temp.)

Pre-set temp. equilibration

4 channels input : in. 2 + ex. 2

Layout of -815

CDME-ray

SH

S1

M0LS

M1

M2

M3

P1M4

M5

P2

S3 L F

S2

PM

O-ray

Modulator for circular polarization

Modulator for circular polarization

Peltier thermostat cell holder

Automated titration unit

Stopped flow attachment

Permanent, electro and super-conducting magnet

FDCD attachment

Etc….

Wide ranges of accessories

Spectra Manager software

JASCO J-815

45 years experience, worldwide leading in

CD spectroscopy

Patented optical system ensure optimal

performance

Best selling CD instrument in the world

(more than 40 units had been sold at Taiwan & HK)

Professional service team

– SUNWAY SCIENTIFIC CO.

JASCO CD User List After 2000

1 高雄醫學大學 藥學系 2000

2 國立台北科技大學 化工系 2000

3 香港理工大學 應用生物及化學系 2000

4 香港中文大學 生化系 2001

5 國立陽明大學 生命科學系 2002

6 國立中山大學 生物醫學研究所 2003

7 國立交通大學 生科系 2003

8 國立台灣大學 化學系 2004

9 國立台灣大學 生化所 2004

10 國立交通大學 應化系 2006

11 中研院 基因體中心 2006

12 國立中興大學 生科系 2006

13 國立嘉義大學 應化系 2006

14 香港科技大學 2006

15 國立台灣大學 化學系 2006

16 國立台灣大學 化工系 2006

17 中研院 化學研究所- 2006

18 陸軍軍官學校 2007

19 文化大學化學系 2007

20 中正大學生科系 2008

References

Kelly, S. M. and Price N. C. (1997) The application of circular dichroism to studies of protein folding and unfolding. Biochim. Biophys. Acta 1338, 161-185.

Sharon M. Kelly, Thomas J. Jess, Nicholas C. Price* (2005) How to study proteins by circular dichroismBiochimica et Biophysica Acta 1751 119 – 139.

Protein structure IRL press

http://www.cryst.bbk.ac.uk/cdweb/html/home.html

SELCON3

CDSSTR

Thanks for your attention!

Daniel Wu

SUNWAY SCIENTIFIC CO.