# Presentation: 888722

## Electrostatic Potential onHLA-DPB1*1701 and HLA-DPB1*0401:Implications for Putative Mechanism of Chronic Beryllium Disease

- Health Effects Laboratory Division,
- National Institute for Occupational Safety and Health,
- Morgantown, WV

- James A. Snyder, Ainsley Weston, Sally S. Tinkle and _Eugene Demchuk_

## Objective

- to understand the physical-chemical reasons for beryllium toxicity
- in Chronic Beryllium Disease (CBD)

## Periodic Table

- What makes Be toxic?

## The Test Hypothesis

- 	Be2+ binds to HLA-DP and changes the specificity of molecular recognition
- Goal:	To identify and test the Be2+ binding 		mode

## Experimental  Evidence

- Amicosante, M., Sanarico, N. Berretta, F., Arroyo, J., Lombardi, G., Lechler, R., Colizzi, V. and Saltini, C.  Beryllium Binding to HLA-DP Molecule Carrying the Marker of Susceptibility to Berylliosis Glutamate 69. *Human Immunology, ***62**, 686-693 (2001)
- Fontenot, A.P., Torres, M., Marshall, W.H., Newman, L.S. and Kotzin, B.L. Beryllium Presentation to CD4+ T Cells Underlies Disease-Susceptibility HLA-DP Alleles in Chronic Beryllium Disease. *Proc. Natl. Acad. Sci. USA,* **97**, 12717-12722 (2000)

**Notes:**
Experimental evidence be binds near glu69 
Saltini, Fentenot

## MHC class II

## Crystal Structure of Human Class II MHC Proteinsin the Protein Data Bank (PDB)

_**HLA-DR1**_
**1AQD**	HLA-DR1 (DRA, DRB1 0101) Human MHC Class II complexed with endogenous peptide
**1DLH**	HLA-DR1 (DRA, DRB1 0101) Human Class II histocompatibility protein (extracellular domain)
**1FYT**	HLA-DR1, DRB1 0101
**1HQR**	DRA*0101 + DRB5*0101
**1KG0**	the Epstein-Barr virus Gp42 Protein bound to the MHC Class II HLA-DR1
**1SEB**	complex of the Human MHC Class II Glycoprotein HLA-DR1 and the bacterial superantigen SEB
_**HLA-DR2**_
**1BX2**	HLA-DR2 (DRA*0101, DRB1*1501) complexed with a peptide from Human myelin basic protein
**1FV1**	immunodominant peptide from myelin basic protein in different registers by two HLA-DR2 alleles
_**HLA-DR3**_
**1A6A**	clip bound to HLA-DR3
_**HLA-DR4**_
**1D5M**	HLA-DR4 complexed with peptide and SEB
**1D5X**	HLA-DR4 complexed with dipeptide mimetic and SEB
**1D5Z**	HLA-DR4 complexed with peptidomimetic and SEB
**1D6E**	HLA-DR4 complex with peptidomimetic and SEB
**1J8H**	HLA-DR1, DRA + HLA-DR4, DRB1*0401
**2SEB**	HLA-DR4 complexed with a peptide from Human collagen II
_**HLA-DQ8**_
**1JK8**	Human insulin peptide-HLA-DQ8 complex
_**HLA-DM**_
**1HDM**	histocompatibility antigen HLA-DM

## Sequence Alignment

**of **_**HLA-DPB1*1701**_** with ******** chain of **_**HLA-DR2**_** (PDB: 1FV1B)**
-                                       10                        20                        30                      40
-   *1701  **RA**T**PENYVH**Q**LRQ**EC**YA**FNGT**Q**R **- -** FL**E**R**Y**IYN**R**EE**FV**RFDS
-   1FV1 **GD**T**RPRFLQ**Q**DKY**EC**HF**FNGT**E**R**VR**FL**H**R**D**IYN**Q**EE**DL**RFDS
-                                      10                       20                       30                     40
-                                      50                       60                       70                        80
-   *1701  DVGE **F**RAVTELGRPD**ED**YWNSQKD**I** LE**EE**RA**VP**D**RM**CRHN
-   1FV1 DVGE**Y**RAVTELGRPD**AE**YWNSQKD**F**LE**DR**RA**AV**D**TY**CRHN
-                                  50                       60                       70                        80
-                                     90                      100                    110                     120
-   *1701  Y**ELD**E**AV**T**L**QRRV**Q**PKV**N**V**S**P**SKKGP**LQHHNLLVC**H**V**TD**F
-   1FV1 Y**GVG**E**SF**T**V**QRRV**E**PKV**T**V**Y**P**ARTQT**LQHHNLLVC**S**V**NG**F
-                                90                     100                     110                     120
-                                   130                     140                   150                      160
-   *1701  YPGSI**Q**VRWF**L**N**G**QEE**T** AGVVST**N**LI**R**NGDWTFQ **I**LVMLE
-   1FV1 YPGSI**E**VRWF**R**N**S** QEE**K**AGVVST**G**LI**Q**NGDWTFQ**T**LVMLE
-                             130                     140                    150                      160
-                                    170                    180                     190                     200
-   *1701  **MT**P**QQ**G**D**VY **I**CQVEH**T**S**LD**SP**V**TVEW**K**A**QSDSAQSKTLTG**
-   1FV1 **TV**P **RS** G**E**VY**T**CQVEH**P**S**VT**SP**L**TVEW**R**A
-                              170                     180                     190

**Sequence Alignment**

## Molecular Dynamics...

## Total Deviation

- at 1 Pico-second intervals

**Total Deviation**

## Total Chargeon  HLA-DP  proteins

- DR Template Protein: 			  -14
- 								   __
- HLA-DPA1*01031/B1*0401: 	-17/-3
- HLA-DPA1*01031/B1*1701: 	-17/-9

- Be2+

## CBD Susceptibility Odds and the Charge on -chain of HLA-DP 

- 	  HLA-DPB1		  Odds	    Charge
- 		*1701					-9
- 		*0601					-9
- 	*0901, *1001				      -9, -9
- 		*1901					-8
- 		*0201					-7
- 		*0401					-3

## Sequence Alignment of HLA-DPB1*1701 with HLA-DPB1*0401

** **            1                                                                                                      40
-     *1701  RATPENY**VH**Q LRQECYAFNG TQRFLERYIY NREEF**V**RFDS
-     *0401  RATPENY **LF**QGRQECYAFNG TQRFLERYIY NREEF**A**RFDS
-                41                                 **-1-1**                                **-2**                           80
-    *1701  DVGEFRAVTE LGRP**DE****D**YWN SQKDILEE**E**RAVPDRMCRHN
-    *0401  DVGEFRAVTE LGRP**AA****E**YWN SQKDILEE**K**RAVPDRMCRHN
-                81   **-1-1**                                                                                          120
-    *1701  YEL**DE****A V**TLQRRVQP**K**VNVS PSKKGPLQHH NLLVCHVTDF
-    *0401  YEL**GG****PM**TLQRRVQP**R**VNVS PSKKGPLQHH NLLVCHVTDF
-                121                                                                                                 160
-    *1701  YPGSIQVRWF LNGQEETAGV VSTNLIRNGD WTFQILVMLE
-    *0401  YPGSIQVRWF LNGQEETAGV VSTNLIRNGD WTFQILVMLE
-                161                                                                 188
-    *1701  MTPQQGDVY **I** CQVEHTSLDS PVTVEWKA
-    *0401  MTPQQGDVY**T** CQVEHTSLDS PVTVEWKA

**Sequence Alignment of HLA-DPB1*1701 with HLA-DPB1*0401**

## -chain

- -chain

- Class II MHC antigen-binding Pocket

- -chain

## Poisson-Boltzmann Equation

## Finite Difference PB Methods

- using divergence theorem:

## HLA-DPA1*01031/B1*0401 (model)

**HLA-DPA1*01031/B1***_**0401**_** (model)**

**Electrostatic Potential Surface Map**

## HLA-DPA1*01031/B1*1701 (model)

**HLA-DPA1*01031/B1***_**1701**_** (model)**

**Electrostatic Potential Surface Map**

## *1701 MEP Movie

## *0401 MEP Movie

## 1701 MEP Movie

## Conclusion

- 	Our results are consistent with the hypothesis that glutamic acid at position 69 of the -chain of HLA-DP may be a determinant of susceptibility to Chronic Beryllium Disease

## People

- _Eugene Demchuk_
- Ainsley Weston		genetics
- Sally S. Tinkle			immunology

**People**

- Health Effects Laboratory Division,
- National Institute for Occupational Safety and Health,
- Morgantown, WV

- computational studies

## Future Goal:  To model interactions of Be2+ with HLA-DP proteins